ES2268780T3 - DETERGENT COMPOSITIONS THAT INCLUDE A MANANASA AND A POLYMER FOR THE RELEASE OF DIRT. - Google Patents

Abstract

A laundry detergent composition comprising a mannanase enzyme, a polymer for the release of dirt for cotton fabrics of the polyethyleneimine type and at least one additional detergent component.

Description

Detergent compositions comprising a Mannanase and a polymer for the release of dirt.

Field of the Invention

The present invention relates to compositions laundry detergents comprising a mannanase and a polymer for the release of dirt in cotton fabrics. This polymer for dirt release is a polyamine modified water soluble and / or dispersible having chain residues main functionalized and better stability against bleach.

Background of the invention

Numerous agents are known in the art for release dirt from use in domestic and industrial processes for the treatment of fabrics such as laundry, drying of fabrics in hot-air clothes dryers, and the like. At market there are different agents to release dirt that are currently used in detergent compositions and articles and fabric softening and antistatic compositions. These polymers for dirt release comprise typically an "main chain" of the oligomeric ester type or polymeric

Until now it has not been possible to develop an effective agent to release dirt in cotton fabrics for use in a laundry detergent. Some attempts to apply the paradigm of formulating the structure of a polymer for the release of dirt imitating the structure of the tissues, a method that was effective in the field of polymers for the release of dirt in polyester fabrics, only they have produced marginal results when it has been applied to agents to release dirt in cotton fabrics. The use of methylcellulose, a cotton polysaccharide with modified oligomeric units, proved to be more effective for polyester than cotton fabrics. For example, in GB-1,314,897, published on April 26, 1973, a hydroxypropyl methylcellulose material is described to prevent redeposition of wet spots and improve the release of dirt in washed fabrics. US Patent 3,897,026, issued to Kearney, describes cellulosic textile materials that have improved soil release and stain resistance properties obtained by reacting an ethylene-maleic anhydride copolymer with the hydroxyl moieties of cotton polymers. . US Patent 3,912,681, issued to Dickson, describes a composition for applying a non-permanent dirt release finish to cotton fabrics comprising a polycarboxylate polymer at a pH lower than 3. US Patent 3,948,838 , granted to Hinton et al ., describes polyacrylic polymers of high molecular weight (500,000-1,500,000) for the release of dirt, preferably used with other tissue treatments. In US Patent 4,559,056, issued to Leigh et al., A process for treating cotton or synthetic fabrics with a composition comprising an elastomeric organopolysiloxane, a copolymeric cross-linking agent of organosiloxanoxyalkylene and a siloxane hardening catalyst is described. . Other agents for releasing dirt that do not comprise terephthalate or mixtures of polyoxy ethylene / propylene are the vinylcaprolactam resins described by Rupert et al. in US-4,579,681 and US-4,614,519. Examples of alkoxylated polyamines and quaternized alkoxylated polyamines suitable for use as dirt dispersants are described in EP-206,513. WO97 / 42288 describes agents for releasing dirt for effective cotton articles that can be prepared from certain modified polyamines available for all cotton articles, whether washed in the presence of a bleaching agent or not. In addition to the aforementioned patents, other patents are mentioned below where different polymers for the release of dirt or modified polyamines are described: US 5,565,145, granted to Watson et al. on October 15, 1996; US 4,548,744, granted to Connor on October 22, 1985; US 4,597,898, granted to Vander Meer on July 1, 1986; US 4,877,896, granted to Maldonado et al. on October 31, 1989; US 4,891,160, granted to Vander Meer on January 2, 1990; US 4,976,879, granted to Maldonado et al. on December 11, 1990; US 5,415,807, issued to Gosselink on May 16, 1995; US 4,235,735, granted to Marco et al. on November 25, 1980; WO 95/32272, published November 30, 1995; GB-1,537,288, published December 29, 1978; GB-1,498,520, published January 18, 1978; DE-28 29 022, granted on January 10, 1980; Japanese unexamined patent application JP 06313271, published on April 27, 1994.

However, the use of these polymers for dirt release in cotton fabrics is not effective enough to protect clothing against stain embedding, in particular cosmetic stains and foods. In fact, cosmetic and food compositions modern increasingly contain additives, such as gums hydrocolloids, which are used as thickeners. In different cosmetic and food compositions are used tomorrow, gum guar and locust bean gum (see Industrial Gum, 2nd edition, R.L. Whistler, p. 308, Academic Press, 1973, ISBN, 0-12-74-6252-x). It is known that these hydrocolloid gums have a very affinity elevated towards cellulosic materials and are difficult to remove. Currently, the use of polymers for the release of dirt in cotton fabrics is not enough to treat these Inlays of cosmetic or food stains.

Food and cosmetic stains and dirt make up the majority of the stains / dirt relevant to the consumer and often comprise food additives such as thickeners or stabilizers. In fact, hydrocolloid gums and emulsifiers are commonly used food additives. The term "gum" represents a group of polysaccharides useful in industry (long chain polymer) or its derivatives that are hydrated in hot or cold water to form viscous solutions, dispersions or gels. The gums are classified as natural and modified. Natural gums include algae extracts, vegetable extrudates, seed or root gums and gums obtained by microbial fermentation. Modified (semi-synthetic) gums include cellulose and starch derivatives and certain synthetic gums such as pectin with low methoxyl content, propylene glycol alginate and carboxymethyl and hydropropyl guar gum (Gums in Encyclopedia Chemical Technology 4th ed. Vol. 12, pp. 842-862, J. Baird, Kelco division of Merck). See also Carbohydrate Chemistry for Food Scientists (Eagan Press - 1997) by RL Whistler and JN BeMiller, chap. 4, p. 63-89 and Direct Food Additives in Fruit Processing by P. Laslo, Bioprinciples and Applications, vol 1, chap. II, p. 313-325 (1996) Technomie publishing. Some of these gums, such as guar gum (E412) or locust bean gum (E410), are generally used alone or combined in numerous food applications (Gums in ECT 4th ed., Vol. 12 pp. 842-862, J. Baird, Kelco division of Merck).

Guar gum in these food and cosmetic products stains comes from the seed endosperm of the Cyamopsis tetragonoloba legume plant. Guar gum (also called guarana) extracted from dicot seed is composed of a main chain with a 1-4 unit, bD-manopyranosyl and is used as a thickener in seasonings and frozen foods and in cosmetic products (H.-D. Belitz , Food Chemistry, pp. 243, English version of the second edition, Springer Verlag, 1987, ISBN 0-387-15043-9 [US]), (Carbohydrate Chemistry for Food Scientists, RL Wilstler, Eagan Press, 1997, ISBN 0 -913250-92-9) e (Industrial Gum, 2nd edition, RL Whistler pp. 308, Academic Press, 1973, ISBN, 0-12-74-6252-x). Locust bean gum (also called San Juan Bautista bread) is also used in the food industry and is extracted from the seed of an evergreen tree that is grown in the Mediterranean area. Locust bean gum probably differs from the structure of guar gum only in that it has a smaller number of D-galactosyl side chains and both have the same main chain of 1-4, bD-manopyranosyl. In leguminous seeds, the water-soluble galactomannan is the main storage carbohydrate and in some cases it comprises up to 20% of the total dry weight. Galactomannan has an α-galactose linked to O-6 of mannose residues and can also be acetylated to different degrees in O-2 and O-3 of the mannose residue.

As described above, follow there is a need to formulate detergent compositions of laundry that provide greater cleaning capacity, especially for cosmetic and food stains, and present advantages of dirt release. This objective has been satisfied formulating laundry detergent compositions comprising a mannanase and a polymer for the release of the dirt in cotton fabrics.

It has also been discovered that the performance of the laundry detergent compositions herein invention is improved by adding other detergent ingredients selected from a detergency builder additive, especially a zeolite, a sodium tripolyphosphate and / or silicate laminar, a surfactant, preferably a nonionic surfactant as alkylethoxylate or alkylmethyl glucamide, a polymer for conventional dirt release and / or mixtures of same.

Mananases have been identified in different Bacillus organisms. For example, Talbot et al., Appl. Environ. Microbiol., Vol. 56, nº 11, p. 3505-3510 (1990) describes a β-mannanase derived from Bacillus stearothermophilus in dimeric form having a PM of 162 kDa and an optimum pH of 5.5-7.5. Mendoza et al., World J. Micobio. Bioech., Vol. 10, No. 5, p. 551-555 (1994) describes a β-mannanase derived from Bacillus subtilisis that has a PM of 38 kDa, an optimal activity at pH 5.0 / 55 ° C and a pI of 4.8. J0304706 describes a β-mannanase derived from Bacillus sp . which has a PM of 37 +/- 3 kDa measured by gel filtration, an optimum pH of 8-10 and a pI of 5.3-5.4. J63056289 describes the production of a thermostable, alkaline β-mannanase that hydrolyzes β-1,4-D-manpiranoside bonds of, for example, mannan and produces hand: oligo: saccharides. Patent J63036774 refers to a Bacillus FERM P-8856 microorganism that produces β-mannanase and β-mannosidase, at an alkaline pH. A purified mannanase from Bacillus amiloliquefaciens and its method of preparation for use in bleaching pulp and paper is described in WO97 / 11164. In WO91 / 18974 a hemicellulase is described as glucanase, xylanase or mannanase, active at extreme pH and temperature as well as the production thereof. WO94 / 25576 describes an enzyme that has a mannanase activity derived from Aspergillus aculeatus CBS 101.43 that can be used for different purposes where degradation or modification of the cell wall of plants or algae is desired. In WO93 / 24622 a mannanase isolated from Trichoderma reesie for bleaching lignocellulosic pastes is described.

However, the synergistic combination of a Mannanase and a polymer for the release of dirt in tissues of cotton to achieve greater cleaning performance and release of dirt in a washing detergent composition of Clothes have never been recognized so far.

Summary of the invention

The present invention relates to compositions laundry detergents comprising a mannanase and polymer for the release of dirt on cotton fabrics that provide greater cleaning performance and release of the dirt.

Detailed description of the invention

An essential element of the composition laundry detergent of the present invention is an enzyme Manana

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The enzyme Mannanase

The present invention encompasses the following three Mannan degradation enzymes: EC 3.2.1.25: β-Mannosidase, EC 3.2.1.78: Endo-1,4- β-mannosidase, Hereafter referred to as "Mannanase" and EC 3.2.1.100: 1,4-?-Manobiosidase (IUPAC Classification- Enzyme nomenclature, 1992 ISBN 0-12-227165-3 Academic Press).

More preferably, the compositions laundry detergents of the present invention comprise a β-1,4-mannosidase (E.C. 3.2.1.78) mentioned as mannanase. The expression "mannanasa" or "galactomannanase" represents a mannanase enzyme defined according to the technique with the official name of manano endo-1,4-beta-mannosidase  and that has the alternative names of beta mannanase and endo-1,4-mannanase and that catalyzes the reaction: random joint hydrolysis 1,4-beta-D-Mannosidic in mannan, galactomannan, glucomannan and galactoglucomannan.

In particular, the mananasas (EC 3.2.1.78) they constitute a group of polysaccharides that degrade mannan and they represent enzymes that are capable of cleaving chains of poliosa which contain mannose units, that is, they are able to cleave glycosidic bonds in mannan, glucomannan, galactomannan and galactogluco-mannan. The mannan are polysaccharides that have a main chain composed of 1,4-mannose bound in 1.4; the glucomannan are polysaccharides that have a main chain or β-mannose and glucose bound in 1.4 form alternating more or less regular; the galactomannans and the galactoglucomannan are mannan and glucomannan with ramifications α-galactose side joined at 1.6. These Compounds can be acetylated.

The degradation of galactomannans and galactoglucomannan is facilitated by a total elimination or partial lateral branches of galactose. Also the degradation of mannan, glucomannan, galactomannan and acetylated galactoglucomannan is facilitated by the total or partial deacetylation. Acetyl groups can be eliminated by alkali or by mannan acetylesterase. Oligomers that are released by mananasas or by a combination of mananasas and α-galactosidase and / or acetyl mannan esterases can be further degraded to release maltose free by β-mannosidase and / or the β-glucosidase.

Mananases have been identified in different Bacillus microorganisms. For example, Talbot et al., Appl. Environ. Microbiol., Vol. 56, nº 11, p. 3505-3510 (1990) describe a beta-mannanase derived from Bacillus stearothermophilus in a dimeric form with a molecular weight of 162 kDa and an optimum pH of 5.5 to 7.5. Mendoza et al., World J. Microbiol. Biotech., Vol. 10, No. 5, p. 551-555 (1994) describes a beta-mannanase derived from Bacillus subtilis having a molecular weight of 38 kDa, an optimal activity at pH 5.0 and 55 ° C and a pI of 4.8. JP-0304706 describes a beta-mannanase derived from Bacillus sp . which has a molecular weight of 373 kDa measured by gel filtration, an optimum pH of 8-10 and a pI of 5.3-5.4. JP-63056289 describes the product of a thermostable alkaline beta-mannanase that hydrolyzes beta-1,4-D-mannopyranoside bonds of, for example, mannans and produces hand-oligosaccharides. JP-63036774 refers to the microorganism Bacillus FERM P-8856 that produces beta-mannose and beta-mannosidase at alkaline pH. JP-08051975 describes alkaline beta-mannanases of Bacillus sp . AM-001 alkalophile. A purified Bacillus amyloliquefaciens mannanase useful for bleaching pulp and paper and a method for preparing them is described in WO 97/11164. WO 91/18974 describes a hemicellulase such as an active glucanase, xylanase or mannanase under extreme conditions of pH and temperature. In WO 94/25576 an enzyme of Aspergillus aculeatus , CBS 101.43 is described which exhibits mannanase activity and can be useful for degrading or modifying the material of the cell wall of plants or algae. In WO 93/24622 a mannanase isolated from Trichoderma reseei useful for bleaching lignocellulosic pastes is described. A hemicellulase capable of degrading a hemicellulose containing mannan is described in WO91 / 18974 and a purified mannanase of Bacillus amiloliquefaciens is described in WO97 / 11164.

In particular, this mannanase enzyme will be an alkaline mannanase as defined below, most preferably a mannanase from a bacterial source. Especially, the laundry detergent composition of the present invention will comprise an alkaline mannanase selected from the mannanase of the Bacillus agaradherens strain and / or of the Bacillus subtilisis 168 strain, yght gene.

The term "alkaline mannanase enzyme" means that it comprises enzymes with an enzymatic activity of at minus 10%, preferably at least 25%, more preferably from at least 40% of its maximum activity at a given pH of 7 to 12, preferably from 7.5 to 10.5.

Most preferably, the laundry detergent composition of the present invention will comprise the alkaline mannanase of Bacillus agaradherens . That manana is

i)

a polypeptide produced by Bacillus agaradherens , NCIMB 40482, or

ii)

a polypeptide comprising an amino acid sequence as shown in positions 32-343 of SEC. 2, or

iii)

a polypeptide analog defined in i) or ii) which is at least 70% homologous with said polypeptide, or derived from said polypeptide by substitution, deletion or addition of one or more amino acids, or is immunologically reactive with an antibody polyclonal directed against said polypeptide in form purified.

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The present invention also encompasses a polypeptide isolated with mannanase activity selected from the group consisting of:

(to)

polynucleotide molecules that encode a polypeptide with mannanase activity and which comprise a nucleotide sequence as shown in SEC. 1 nucleotide 97 to nucleotide 1029;

(b)

homologous species of (a);

(C)

polynucleotide molecules that encode a polypeptide with mannanase activity that is identical to 70% less than the amino acid sequence SEC. 2 from amino acid residue 32 to amino acid residue 343;

(d)

molecules complementary to (a), (b) or (C); Y

(and)

degenerate nucleotide sequences of (a), (b), (c) or (d).

Plasmid pSJ1678 comprising the polynucleotide molecule (the DNA sequence) encoding a mannanase of the present invention has been transformed into an Escherichia coli strain that has been deposited by the inventors, in accordance with the Budapest Treaty on Recognition International Deposit of Microorganisms for the Purposes of Patent Procedure, at Deutsche Sammlung von Mikroorganismen und Zellkulturen GmbH, Mascheroder Weg 1b, D-38124 Braunschweig, Federal Republic of Germany, on May 18, 1998 with the deposit number DSM 12180.

A second highly preferred enzyme is the mannanase of strain 168 of Bacillus subtilisis , where the mannanase:

i)

 is encoded by the coding part of the DNA sequence that is shown in SEC. 5 or an analogue of said sequence and / or

ii)

a polypeptide comprising an amino acid sequence according to SEC. 6 or

iii)

a polypeptide analog defined in ii) which is homologous at least in 70% to said polypeptide, or is derived from said polypeptide by substitution, deletion or addition of one or more amino acids, or reacts immunologically with a polyclonal antibody against said polypeptide in purified form.

The present invention also encompasses a polypeptide isolated with mannanase activity selected from the group consisting of:

(to)

polynucleotide molecules that encode a polypeptide with mannanase activity and which comprise a nucleotide sequence according to SEC. 5

(b)

homologous species of (a);

(C)

polynucleotide molecules that encode a polypeptide with mannanase activity that is identical to 70% less than the amino acid sequence SEC. 6)

(d)

molecules complementary to (a), (b) or (C); Y

(and)

degenerate nucleotide sequences of (a), (b), (c) or (d).

Definitions

Before studying the present invention in more detail the following terms must be defined:

The term "ortholog" (or "species homologous ") means a polypeptide or a protein obtained from a species that has homology with a polypeptide or protein analogue of a different species.

The term "paralogue" means a polypeptide or a protein obtained from a certain species that has homology with a polypeptide or a different protein from the same species

The term "expression vector" means a DNA molecule, linear or circular, comprising a segment which encodes a polypeptide of interest operably linked to additional segments that perform their transcription. These segments Additional may include promoter and terminator sequences and may optionally include one or more sources of replication, one or more selectable markers, an enhancer, a signal of polyadenylation, and the like. Expression vectors are derived generally of plasmid or viral DNA or may contain elements of both of them. The expression vector can be any vector of expression conveniently subjected to DNA procedures recombinant and the choice of vector will often depend on the host cell into which the vector must be introduced. So, the vector it can be a vector that replicates autonomously, that is a vector that exists as an independent chromosomal entity whose replication is independent of chromosomal replication, e.g. eg a plasmid Alternatively, the vector may be one that, when introduced into a host cell, it integrates into the genome of the host cell and replicates together with the chromosome (s) in which it has been integrated.

The term "expressed in a manner recombinant "used herein in relation to the expression of a polypeptide or protein follows the standard definition in the technique Recombinant expression of a protein is performed generally using an expression vector as described previously.

The term "isolated", when applied to a polynucleotide molecule, means that the polynucleotide has been taken from its natural genetic environment and therefore is free of other strange or unwanted coding sequences and is found in a form suitable for use in production systems of genetic engineering proteins These isolated molecules are separated from their natural environment and include cDNA and genomic clones. The isolated DNA molecules of the present invention are free of other genes with which they are normally associated but may include untranslated 5 'and 3' natural regions such as promoters and terminators. The identification of associated regions It is evident to the person skilled in the art (see, for example, Dynan and Tijan, Nature 316: 774-78, 1985).

The term "an isolated polynucleotide" they can also be replaced by "a cloned polynucleotide". When the term "isolated" applies to a protein / polypeptide indicates that the protein is in a condition different from its natural environment. In a preferred form, the Isolated protein is virtually free of other proteins, especially from other homologous proteins (ie "impurities counterparts "[see below]). It is preferable to provide the protein in a pure form greater than 40%, more preferably in a pure form greater than 60%. Even more preferably it is preferred provide the protein in a high pure form, that is, with a pure greater than 80%, more preferably greater than 95% and still more preferably greater than 99%, according to the determination SDS-PAGE.

The term "isolated protein / polypeptide" it can also be substituted by "protein / polypeptide purified. "

The term "homologous impurities" means any impurity (e.g., another polypeptide other than polypeptide of the invention) that comes from the homologous cell of the that the polypeptide of the invention was originally obtained. He term "obtained from" used herein in relationship with a specific microbial source means that the polynucleotide and / or the polypeptide comes from a specific source or of a cell in which a source gene has been inserted.

The term "operably linked", referred to DNA segments, it means that the segments are arranged so that they work according to their purposes provided, p. eg, that transcription be initiated in the promoter and proceed through the coding segment to the terminator.

The term "polynucleotide" means a single or double stranded base deoxyribonucleotide or ribonucleotide polymer read from the 5 'end to the 3' end. The polynucleotides include RNA and DNA and can be isolated from natural sources, synthesized in vitro or prepared from a combination of natural and synthetic molecules.

The term "molecule complements of polynucleotide "means polynucleotide molecules that have a complementary base sequence and an inverted orientation with respect to a reference sequence. For example, the sequence 5 'ATGCACGGG 3' is complementary to the 5 'CCCGTGCAT 3'.

The term "degenerate sequence of nucleotides "means a nucleotide sequence that includes one or more degenerate codons (with respect to a molecule of reference polynucleotide encoding a polypeptide). The degenerate codons contain different nucleotide triplets but encodes the same amino acid residue (i.e. GAU and GAC triplets each code Asp).

The term "promoter" means a portion of a gene that contains DNA sequences that bind RNA polymerase and begin transcription. The promoter sequences are they find normally, but not always, in regions not 5 'gene encoders.

The term "signal sequence of secretion "means a DNA sequence that encodes a polypeptide (a "secretion peptide") that, as a component of a major polypeptide, directs the major polypeptide through a secretion pathway of a cell in which it is synthesized. Generally the major peptide is cleaved to remove the peptide of secretion during transit through the route of secretion.

How to use a sequence of the invention to obtain others related sequences

The information included in this report relative to a polynucleotide sequence encoding a Mannanase of the invention can be used as a tool for identify other homologous mananasas. For example, it can be used polymerase chain reaction (PCR) to amplify sequences encoding other homologous mannanases from different microbial sources, in particular of different species of Bacillus.

Activity test assessment

The mannanase activity of a polypeptide of the invention with mannanase activity can be analyzed by methods standard test known in the art, such as the application of an experimental solution to 4 mm diameter holes made on agar plates containing 0.2% AZCL galactomannan (locust bean gum), that is, a substrate for essay of endo-1,4-beta-D-mannanase available as CatNo. I- AZGMA of the company Megazyme, 110.00 dollars / 3 grams (Megazyme website: http://www.megazyme.com/Purchase/index.html).

Polynucleotides

An isolated polynucleotide of the invention is hybridizes to regions of similar size of the SEC. 1, or to a sequence complementary to this, under conditions of medium stringency such as minimum.

In particular the polynucleotides of the invention hybridize to a double stranded DNA probe denatured comprising the complete sequence shown in positions 97-1029 of SEC. 1 or any probe comprising a sub sequence of SEC. 1 with a length of at least about 100 base pairs under conditions of average stringency at least, but preferably in conditions High stringency as described in detail below. The adequate experimental conditions to determine hybridization medium to high stringency between a nucleotide probe and a homologous DNA or RNA sequence involves prior wetting of the filter that contains the DNA or RNA fragments that are going to hybridize in 5 x SSC (sodium chloride / sodium citrate, Sambrook et al. 1989) for 10 min and the previous hybridization of the filter in a 5 x SSC solution, 5 x Denhardt solution (Sambrook et al. 1989), 0.5% SDS and 100 µg / ml salmon sperm DNA denatured sonicated (Sambrook et al. 1989), followed by hybridization in the same solution that contains a concentration of 10 ng / ml of a random barley probe (Feinberg, A. P. and Vogelestein, B. (1983) Anal. Biochem 132: 6-13), labeled 32P-dCTP (higher specific activity at 1 x 109 cpm / \ mug) for 12 hours at approx. 45 ° C Then the filter is washed twice for 30 minutes in 2 x SSC, SDS at 0.5% at least at 60 ° C (medium stringency), even more preferably at less at 65 ° C (medium / high stringency), even more preferably at less at 70 ° C (high stringency) and even more preferably at least at 75ºC (very high stringency).

The molecules to which the probe hybridizes oligonucleotides under these conditions are detected with a film X-ray.

As mentioned above, the Isolated polynucleotides of the present invention include DNA and RNA The methods for isolating DNA and RNA are well known in the technique. The genes encoding DNA and RNA of interest can be cloned in libraries or DNA libraries using methods known in the art.

The following are identified and isolated polynucleotides encoding polypeptides with mannanase activity of the invention, for example, by hybridization or PCR.

The present invention also provides counterpart polypeptides and polynucleotides of different bacterial strains (orthologs or paralogs). Of special interest are the polypeptides with mannanase activity of gram-positive alkalophilic strains, including the Bacillus species.

Homologous species of a mannanase activity polypeptide of the present invention can be cloned using the information and compositions provided by the present invention together with conventional cloning techniques. For example, a DNA sequence of the present invention can be cloned using chromosomal DNA obtained from a type of cell that expresses the protein. Suitable sources of DNA can be identified by analyzing Northern blots with probes designed in accordance with the sequences described herein. A library is then prepared from chromosomal DNA of a positive cell line. A DNA sequence of the invention encoding a polypeptide with mannanase activity can be isolated by different methods such as, for example, with probes designed from the sequences described in the present specification and claims or with one or more sets of degenerate probes based on The sequences described. A DNA sequence of the invention can also be cloned using the polymerase chain reaction (PCR) (Mullis, US Pat. No. 4,683,202) using primers designed from the sequences described herein. In another method the DNA library can be used to transform or transfect host cells and the expression of the DNA of interest can be detected with an antibody (monoclonal or polyclonal) directed against the cloned mannanase of B. agaradherens , NCIMB 40482, expressed and purified as described in Materials and Methods and in Example 1, or by an activity assay with a polypeptide with mannanase activity.

The mannanase encoding part of the DNA sequence cloned in plasmid pSJ1678 present in Escherichia coli DSM 12180 and / or an analogous DNA sequence of the invention can be cloned from a strain of the bacterial species Bacillus agaradherens , preferably the strain NCIMB 40482, to obtain the enzyme with degradation activity of mannan or a different or related microorganism as described herein.

Alternatively, the analogous sequence may be constructed from the DNA sequence obtainable from the plasmid present in Escherichia coli DSM 12180 (believed to be identical to the attached SEC. 1), p. eg, of a sub-sequence thereof, and / or by introducing nucleotide substitutions that do not give rise to another amino acid sequence of the mannanase encoded by the DNA sequence, but that corresponds to the codon use of the host microorganism intended for production of the enzyme, or by introducing nucleotide substitutions that may result in a different amino acid sequence (i.e., a variant of the mannan degradation enzyme of the invention).

Polypeptides

Amino acid sequence SEC 32-343 2 is a mannanase sequence mature

The present invention also provides polypeptides with mannanase activity that are practically homologous to the SEC polypeptide. 2 and homologous species (paralogs u orthologs) of them. The term "practically homologous" is used herein to designate polypeptides that they have a sequence identity of 70%, preferably at least 80%, more preferably at least 85% and even more preferably at least 90%, with the sequence of the amino acids 32-343 of SEC. 2 or its orthologs or paralogs These polypeptides will be more preferably identical to at least 95%, and most preferably identical at 98% or more, to amino acid sequence 32-343 of SEC. 2 or its orthologs or paralogs The percentage of sequence identity is determined by conventional methods, using programs computer skills known in the art such as GAP which is included in the GCG program package (Program Manual for the Wisconsin Package, Version 8, August 1994, Genetics Computer Group, 575 Science Drive, Madison, Wisconsin, USA 53711) according to Needleman, S.B. and Wunsch, C.D., (1970), Journal of Molecular Biology, 48, 443-453. For sequence comparison of Polypeptides using the GAP method with the following values: GAP creation penalty 3.0 and GAP extension penalty of 0.1.

The sequence identity of molecules of polynucleotide is determined by similar methods using The following GAP values to compare DNA sequences: GAP creation penalty of 5.0 and GAP extension penalty of 0.3.

The preparation of the enzyme of the invention preferably comes from a microorganism, preferably from a bacterium, an archea or a fungus, especially from a bacterium such as, e.g. eg, a bacterium belonging to Bacillus , preferably from an alkalophilic strain of Bacillus that can be selected from the group consisting of the Bacillus agaradherens species and very related Bacillus species in which all species are preferably at least 95%, and even more preferably at least 98%, homologous to Bacillus agaradherens in terms of aligned 16S rDNA sequences.

Virtually homologous proteins and polypeptides are characterized by having one or more amino acid substitutions, deletions or additions. These changes are preferably of a minor nature, that is, conservative amino acid substitutions (see Table 2) and other substitutions that do not significantly affect the folding or activity of the protein or polypeptide; small deletions, typically from one to about 30 amino acids, and small extensions of amino or carboxyl terminal, such as a methionine residue with amino terminal, a small linker peptide of up to about 20-25 residues or a small extension that facilitates purification (an affinity tag), such as a poly-histidine tract, protein A (Nilsson et al., EMBO J. 4 : 1075, 1985; Nilsson et al., Methods Enzymol . 198 : 3, 1991. See, in general , Ford et al., Protein Expression and Purification 2 : 95-107, 1991. DNA encoding affinity tags are available (eg, Pharmacia Biotech, Piscataway, NJ; New England Biolabs, Beverly, MA).

However, although the changes described previously preferably they are of a minor nature, these changes they can also be of a greater nature such as, e.g. eg, the merger of polypeptides larger than 300 amino acids or more as an extension from amino or carboxyl terminals to a mannanase polypeptide of the invention.

TABLE 1 Amino acid substitutions conservative

Basic

: {} \ arginine, lysine, histidine

Acidic

: {} \ hskip0.5cm glutamic acid, acid aspartic

 Polar

: {} \ hskip0.5cm glutamine, asparagine

Hydrophobes

: {} \ hskip0.5cm leucine, isoleucine, valine

Aromatic

: {} \ phenylalanine, tryptophan, tyrosine

Little

: {} \ wskip0.5cm glycine, alanine, serine, threonine, methionine

In addition to the conventional 20 amino acids, unconventional amino acids (such as 4-hydroxyproline, 6-N-methyl lysine, 2-aminoisobutyric acid, isovaline and a-methyl serine) can also be substituted for amino acid residues of a polypeptide. A limited number of non-conservative amino acids, amino acids that are not encoded by the genetic code and unnatural amino acids can be substituted for amino acid residues. The "unnatural amino acids" have been modified after protein synthesis and / or have a chemical structure in their chain (s)
lateral (s) different from standard amino acids. Unnatural amino acids can be obtained by chemical synthesis, or preferably, are commercial products that include pipecolic acid, thiazolidinecarboxylic acid, dehydroproline, 3-methylproline and 4-methylproline and 3,3-dimethylproline.

Essential amino acids in polypeptides with mannanase activity can be identified by methods known in the art, such as site-directed mutagenesis or alanine detection mutagenesis (Cunningham and Wells, Science 244 : 1081-1085, 1989). In the latter technique, individual alanine mutations are introduced into each residue in the molecule and the resulting mutant molecules analyze biological activity (i.e., mannanase activity) to identify amino acid residues that are critical to the activity of the molecule. See also, Hilton et al., J. Biol. Chem . 271 : 4699-4708, 1996. The active site of the enzyme or other biological interaction can also be determined by analysis of the physical structure with techniques such as, e.g. eg, nuclear magnetic resonance, crystallography, electron diffraction or photoaffinity labeling, along with the mutation of new amino acid contact sites. See, for example, de Vos et al., Science 255 : 306-312, 1992; Smith et al., J. Mol. Biol 224 : 899-904, 1992; Wlodaver et al., FEBS Lett . 309 : 59-64, 1992. The identities of the essential amino acids can also be inferred from the analysis of homologies with polypeptides that are related to a polypeptide according to the invention.

Multiple amino acid substitutions can be made and analyzed using known methods of mutagenesis, recombination and / or redistribution followed by a suitable detection procedure such as those described by Reidhaar-Olson and Sauer ( Science 241 : 53-57, 1988), Bowie and Sauer ( Proc. Natl. Acad. Sci . USA 86 : 2152-2156, 1989), WO95 / 17413, or WO 95/22625. These authors describe methods to simultaneously randomize two or more positions in a polypeptide or recombine / redistribute different mutations (WO95 / 17413, WO95 / 22625) to then select a functional polypeptide and sequence the mutagenized polypeptides and determine the spectrum of permitted substitutions in each position. Other methods that can be used include phage display (e.g., Lowman et al., Biochem . 30 : 10832-10837, 1991; Ladner et al., US-5,223,409; Huse, WIPO Publication WO 92/06204 ) and region-directed mutagenesis (Derbyshire et al., Gene 46 : 145, 1986; Ner et al., DNA 7 : 127, 1988).

The mutagenesis / redistribution methods described above can be combined with detection methods High performance automatic to detect the activity of mutagenized polypeptides cloned into host cells. The mutagenized DNA molecules encoding active polypeptides can be recovered from host cells and sequenced Quickly using modern equipment. These methods allow quickly determine the importance of individual waste of amino acid in a polypeptide of interest and a polypeptides of unknown structure.

Using the mentioned methods previously, the person skilled in the art can identify and / or prepare different polypeptides that are practically homologous to residues 32 to 343 of SEC. 2 and conserve the mannanase activity of the wild protein

Protein Obtaining

Proteins and polypeptides, including full-length proteins, fragments thereof and fusion proteins, can be obtained in genetically engineered host cells by conventional techniques. Suitable host cells are those cell types that can be transformed or transfected with exogenous and cultured DNA and include bacteria, fungal cells and cultured higher eukaryotic cells. Bacterial cells, especially cultured cells of gram-positive microorganisms, are preferred. Gram-positive cells of the genus Bacillus are especially preferred as, e.g. eg., those of the group consisting of Bacillus subtilis, Bacillus lentus, Bacillus brevis, Bacillus stearothermophilus, Bacillus alqualophilus, Bacillus amyloliquefaciens, Bacillus coagulans, Bacillus circulans, Bacillus lautus, Bacillus thuringiensis, Bacillus licheniformis, and Bacillus agaradherens, especially the Bacillus agaradherens .

Techniques for manipulating cloned DNA molecules and introducing exogenous DNA into different host cells have been described by Sambrook et al., Molecular Cloning: A Laboratory Manual , 2nd ed., Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY, 1989; Ausubel et al. (eds.), Current Protocols in Molecular Biology , John Wiley and Sons, Inc., NY, 1987; and " Bacillus subtilis and Other Gram-Positive Bacteria", Sonensheim et al., 1993, American Society for Microbiology, Washington DC

In general, a DNA sequence that encodes a mannanase of the present invention is operably linked to other genetic elements necessary for its expression, which generally includes a promoter and a transcription terminator Within an expression vector. The vector will also contain generally one or more selectable markers and one or more origins of replication, although the person skilled in the art will recognize that in certain systems markers can be provided selectable in separate vectors and that DNA replication exogenous can be performed by integrating into the genome of the host cell The selection of promoters, terminators, markers Selectable, vectors and other elements is a matter of design Routine for the person skilled in the art. Many of these elements They are described in the bibliography and are marketed by market suppliers.

To direct a polypeptide to the secretion pathway of a host cell, a sequence of secretion signals (also known as leader sequence, prepro sequence or presequence) is found in the expression vector. The sequence of secretion signals may be that of the polypeptide, or it may be derived from another secreted or synthesized de novo protein. Numerous suitable secretion signal sequences are known in the art, citing " Bacillus subtilis and Other Gram-Positive Bacteria", Sonensheim et al., 1993, American Society for Microbiology, Washington DC; and Cutting, SM (eds.) "Molecular Biological Methods for Bacillus", John Wiley and Sons, 1990, for a more detailed description of suitable secretion signal sequences, especially for secretion in a Bacillus host cell. The sequence of secretion signals binds to the DNA sequence in the correct reading frame. The secretion signal sequences are generally in the 5 'position with respect to the DNA sequence encoding the polypeptide of interest, although certain signal sequences may be in another position in the DNA sequence of interest (see, e.g. , Welch et al., US 5,037,743; Holland et al., US 5,143,830).

The transformed host cells or transfected are grown by conventional procedures in a culture medium that contains nutrients and other components necessary for the growth of the host cells chosen. In different suitable means are known in the art, including defined media and complex media, and generally include a source of carbon, a source of nitrogen, amino acids Essentials, vitamin and minerals. Media can also contain these components as growth factors or serum, according to the needs The growth medium will generally select cells containing the DNA added exogenously by, by example, selection of the substance or deficiency of a nutrient essential that is complemented by the selectable marker transported in the expression vector or co-transfected in the host cell.

Protein isolation

When the expressed recombinant polypeptide is secreted, this can be purified from the culture medium. Preferably the expression host cells are removed from the means before purification of the polypeptide (for example by centrifugation)

When the expressed recombinant polypeptide does not is secreted from the host cell, the host cell is preferably altered and the polypeptide is released at a aqueous "extract" which is the first phase of these techniques of purification. Preferably the expression host cells are collected from the media before cell disruption (for example by centrifugation).

Cellular alteration can be performed by conventional techniques such as lysozyme digestion or forcing the cells through high pressure. For one more description detailed information on these cell alteration techniques, see Robert K. Scobes, Protein Purification, 2nd edition, Springer-Verlag

Whether or not the polypeptides are secreted recombinants (or chimeric polypeptides) expressed, these may be purified by means and methods of fractionation and / or conventional purification.

To fractionate the samples, the precipitation with ammonium sulfate and extraction with an acid or a chaotropic agent The illustrative purification steps can include hydroxyapatite, size exclusion, FPLC and chromatography High resolution liquid in reverse phase. The means of Suitable anion exchange include derivatized dextrans, agarose, cellulose, polyacrylamide, special silicas, and the like. The derivatives of PEI, DEAE, QAE and Q are preferred, DEAE being Fast-Flow Sepharose (Pharmacia, Piscataway, NJ) especially preferred. Illustrative chromatographic media include the means derivatized with phenyl, butyl or octyl, such as Phenyl-Sepharose FF (Pharmacia), Toyopearl butyl 650 (Toso Haas, Montgomeryville, PA), Octyl-Sepharose (Pharmacia) and the like; or resins polyacrylics, such as Amberchrom CG 71 (Toso Haas) and the like. Suitable solid supports include glass beads, resins based on silica, cellulosic resins, agarose beads, pearls crosslinked agarose, polystyrene beads, resins cross-linked polyacrylamide, and the like, which are insoluble in the conditions in which they are used. These supports can be modified with reactive groups that allow protein binding by amino groups, carboxyl groups, sulfhydryl groups, groups hydroxyl and / or carbohydrate moieties. Coupling Examples include the activation of cyanogen bromide, the activation of N-hydroxysuccinimide, epoxy activation, sulfhydryl activation, hydrazide activation and carboxyl derivatives and amino derivatives for coupling chemistry carbodiimide These media and other solid media are well known. and widely used in the art and are products of commercial suppliers

The choice of a certain method is a matter of routine design and is determined in part by the properties of the chosen support. See, for example, Affinity Chromatography: Principles & Methods , Pharmacia LKB Biotechnology, Uppsala, Sweden, 1988.

The polypeptides of the invention or the fragments thereof can also be prepared by synthesis chemistry. The polypeptides of the invention can be monomers or multimers; glycosylated or non-glycosylated; pegylated or not pegylated; and may or may not include an initial amino acid residue methionine

Using the sequence information described a DNA sequence of full length encoding a mannanase and comprising the DNA sequence shown in SEC. 1, at least the DNA sequence from position 97 to position 1029.

Cloning is done by procedures standards known in the art such as

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prepare a library of a Bacillus strain, especially strain B. agaradherens , NCIMB 40482;

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  place this library in suitable substrate plates;

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  identify a clone that comprises a polynucleotide sequence of the invention by conventional hybridization techniques with a probe based on SEC. one; or

\ blacksquare

Identify a clone of said Bacillus agaradherens NCIMB 40482 library by a reverse PCR strategy using primers based on sequence information from the SEC. 1. Reference is made to MJ MCPherson et al. ("PCR A practical approach" Information Press Ltd, Oxford, England).

In view of the sequence information described herein (SEC. 1, SEC. 2), the isolation of homologous polynucleotide sequences encoding the homologous mannanase of the invention by a similar strategy using libraries of related microbial microorganisms, in Particular of libraries of other strains of the genus Bacillus such as the alkalophilic species of Bacillus , represents a routine work for the person skilled in the art.

Alternatively, the DNA encoding the enzyme of the invention that degrades mannan or galactomannan can be suitably cloned, according to well-known procedures, from a suitable source such as any of the aforementioned microorganisms using synthetic oligonucleotide probes prepared.
strands according to the DNA sequence that can be obtained from the plasmid present in Escherichia coli DSM 12180.

In the same way, the polynucleotide molecule of the invention can be isolated from Escherichia coli , DSM 12180, in which the plasmid obtained by cloning is deposited as described above. Also, the pre
This invention refers to a practically pure isolated biological culture of the Escherichia coli strain, DSM 12180.

In this context the term "preparation of the enzyme "means an enzymatic fermentation product conventional, possibly isolated and purified, from a only species of a microorganism, wherein said preparation normally it comprises different enzymatic activities; or one mixture of monocomponent enzymes, preferably derived enzymes of bacterial or fungal species using recombinant techniques conventional, whose enzymes have been fermented and possibly isolated and purified separately and that may come from different species, preferably fungal species or bacterial; or the fermentation product of a microorganism that acts as a host cell to express a mannanase recombinant, but where said microorganism produces the same other enzymes time, p. e.g., pectin degradation enzymes, proteases or cellulases, being natural fermentation products of the microorganism, that is, the enzyme complex obtained from conventional form by the corresponding microorganism natural.

A method for preparing the Enzyme is the method that involves cultivating a microorganism, e.g. ex. a wild type strain, capable of producing mannanase in conditions that allow to produce the enzyme and recover the enzyme of the crop Cultivation can be performed using techniques of conventional fermentation, p. e.g., culture in jars or fermenters with stirring to ensure sufficient aeration in a growth medium that induces the production of the enzyme Manana The growth medium may contain a source of N conventional as, p. eg, peptone, yeast extract or casamino acids, a reduced amount of a conventional source of C as, p. eg, dextrose or sucrose and an inducer such as guar gum or Locust bean gum. Recovery can be done using conventional techniques such as, e.g. e.g., biomass separation and supernatant by centrifugation or filtration, recovery of supernatant or alteration of cells if the enzyme of interest is intracellular, perhaps followed by another purification according to EP 0 406 314 or by crystallization according to WO 97/15660.

Immunological cross reactivity

Polyclonal antibodies used to determine immunological cross reactivity can be prepared using a purified mannanase enzyme. More specifically, antiserum against the mannanase of the invention can be obtained by immunizing rabbits (or other rodents) according to the method described by N. Axelsen et al. in: A Manual of Quantitative Immunoelectrophoresis, Blackwell Scientific Publications, 1973, chap. 23, or A. Johnstone and R. Thorpe, Immunochemistry in Practice, Blackwell Scientific Publications, 1982 (more specifically on pages 27-31). The purified immunoglobulins can be obtained from the antisera, for example by saline precipitation ((NH4) 2 SO4), followed by dialysis and ion exchange chromatography, for example in DEAE-Sephadex. Immunochemical characterization of proteins can be performed by double diffusion analysis of Outcherlony (O. Ouchterlony in: Handbook of Experimental Immunology (DM Weir, Ed.), Blackwell Scientific Publications, 1967, pp. 655-706), by cross immunoelectrophoresis (N Axelsen et al., Supra , caps 3 and 4), or by rocket-type immunoelectrophoresis (N. Axelsen et al., Chap. 2).

Examples of bacteria useful for obtaining the enzyme or the preparation of the enzyme of the invention are gram-positive bacteria, preferably of the Bacillus / Lactobacillus subdivision, preferably a strain of the genus.
Bacillus , more preferably a Bacillus agaradherens strain, especially the Bacillus agaradherens strain, NCIMB 40482.

Also included is an isolated mannanase that It has the properties described above and is exempt from homologous impurities, and that is obtained using techniques conventional recombinants.

Determination of the catalytic activity (ManU) of the morning

Colorimetric determination: Substrate: 0.2% of AZCL-galactomannan (Megazyme ™, Australia) rubber carob in 0.1 M glycine buffer, pH10.0. The determination is performs in a 1.5 ml Eppendorf microtube in a thermomixer with stirring and with temperature control at 40 ° C. 0.750 were incubated ml of substrate with 0.05 ml of enzyme for 20 min and centrifuged for 4 minutes at 1570.8 rad / s (15000 rpm). The color of the supernatant at 600 nm in a 1 cm cuvette. One ManU (units Mannanase) provides 0.24 abs in 1 cm.

Obtaining Bacillus Agaradherens mananasa NCIMB 40482 Strains

Bacillus agaradherens NCIMB 40482 comprises the enzyme mannanase that encodes the DNA sequence.

E. coli strain: E. coli SJ2 cells (Diderichsen, B., Wedsted, U., Hedegaard, L., Jensen, BR, Sjøholm, C. [1990] Cloning of aldB, which encodes alpha-acetolactate decarboxylase were prepared , an exoenzyme from Bacillus brevis . J. Bacteriol., 172, 4315-4321) and were transformed by electroporation using a BIO-RAD Gene Pulser ™ electroporator according to the supplier's instructions.

B. subtilis PL2306. This strain is B. subtilis DN1885 with altered apr and npr genes (Diderichsen, B., Wedsted, U., Hedegaard, L., Jensen, BR, Sjøholm, C. [1990] Cloning of aldB, which encodes alpha-acetolactate decarboxylase, an exoenzyme from Bacillus brevis . J. Bacteriol., 172, 4315-4321) that is altered in the transcriptional unit of the known cellulase gene of Bacillus subtilis to obtain negative cellulase cells. The alteration was basically performed according to (Eds. AL Sonenshein, JA Hoch and Richard Losick [1993] Bacillus subtilis and other Gram-Positive Bacteria, American Society for microbiology, p. 618).

Competent cells were prepared and transformed as described by Yasbin, RE, Wilson, GA and Young, FE (1975) Transformation and transfection in lysogenic strains of Bacillus subtilis : evidence for selective induction of prophage in competent cells. J. Bacteriol, 121: 296-304.

Plasmids

pSJ1678 (as described in detail in WO 94/19454).

pMOL944: This plasmid is a derivative of pUB110 that basically contains elements that make the plasmid propagable in Bacillus subtilis , kanamycin resistance gene, and that has a strong promoter and signal peptide cloned from the amyL gene of B. licheniformis ATCC14580. The signal peptide contains a SacII site that makes it suitable for cloning the DNA encoding the mature part of a protein infusion with the signal peptide. This results in the expression of a Pre-protein that is directed outside the cell.

The plasmid was constructed using techniques of conventional genetic engineering that are briefly described to continuation.

Construction of pMOL944

Plasmid pUB110 (McKenzie, T. et al., 1986, Plasmid 15: 93-103) was digested with the enzyme of unique restriction NciI. An amplified PCR fragment of the promoter amyL encoded in plasmid pDN1981 (P.L. Jørgensen et al., 1990, Gene, 96, p. 37-41) was digested with NciI e piece of inserted into the NUBI digested pUB110 to obtain the plasmid pSJ2624.

The two PCR primers used have the following sequences:

one

Primer # LWN5494 inserts a NotI site into the plasmid

Then the plasmid pSJ2624 was digested with SacI and NotI and a new PCR fragment amplified in the promoter amyL encoded in pDN1981 was digested with SacI and NotI and this DNA fragment was inserted into the pSJ2624 digested with SacI-NotI to obtain plasmid pSJ2670.

This cloning replaces the first cloning of the amyL promoter with the same promoter but in the direction contrary. The two primers used for PCR amplification They have the following sequences:

2

Plasmid pSJ2670 was digested with enzymes restriction PstI and BclI and an amplified PCR fragment from of a cloned DNA sequence encoding alkaline amylase SP722 (described in the published international patent application as WO95 / 26397) was digested with PstI and BclI and inserted to obtain plasmid pMOL944. The two primers used for the PCR amplification have the following sequence:

3

Cloning of the Bacillus agaradherens mannanase gene Genomic DNA Preparation

The Bacillus agaradherens NCIMB 40482 strain was propagated in liquid medium as described in WO94 / 01532. After 16 hours of incubation at 30 ° C and 31.4 rad / s (300 rpm), the cells were harvested and the genomic DNA isolated by the method described by Pitcher et al. (Pitcher, DG, Saunders, NA, Owen, RJ (1989) Rapid extraction of bacterial genomic DNA with guanidium thiocyanate Lett. Appl. Microbiol., 8, 151-156).

Construction of the library

Genomic DNA was partially digested with the restriction enzyme Sau3A and fractionated by size 0.7% agarose gel electrophoresis. Fragments of 2 a were isolated 7 kb in size by electrophoresis on DEAE cellulose paper (Dretzen, G., Bellard, M., Sassone-Corsi, P., Chambon, P. [1981] A reliable method for the recovery of DNA fragments from agarose and acrylamide gels. Anal. Biochem., 112, 295-298).

The isolated DNA fragments were ligated to the plasmid DNA pSJ1678 digested with BamHI and the ligated mixture was used to transform E. coli SJ2.

Identification of positive clones

A DNA library in E. coli , constructed as described above, was analyzed on LB agar plates containing 0.2% AZCL-galactomannan (Megazyme) and 9 µg / ml chloramphenicol and incubated overnight at 37 ° C. Clones expressing mannanase activity appeared with a blue diffusion halo. Plasmid DNA from one of these clones was isolated with Qiagen plasmid spin preps in 1 ml of overnight broth (cells incubated at 37 ° C in TY with 9 µg / ml chloramphenicol and stirring at 26.2 rad / s [250 rpm]).

This clone (MB525) was also characterized by DNA sequencing of the cloned Sau3A DNA fragment. The DNA sequencing was performed using "firstwalking", using the Taq deoxy-C sequencing kit terminalycle (Perkin-Elmer, USA), terminators with fluorescent label and suitable oligonucleotides as primers

Sequence data analysis was performed. according to Devereux et al. (1984) Nucleic Acids Res. 12, 387-395. The sequence encoding the mannanase is shown in SEC. 1. The protein derived sequence is shown in the SEC. 2.

Subcloning and expression of mannanase in B.subtilis

The mannanase encoding the DNA sequence was PCR amplified using the PCR primer set consisting in these two oligonucleotides:

4

SacII restriction sites underlined and NotII.

The isolated chromosomal DNA of B. agaradherens NCIMB 40482 described above was used as a template in a PCR reaction using Amplitaq DNA Polymerase (Perkin Elmer) according to the manufacturer's instructions. The PCR reaction was performed in PCR buffer (10 mM Tris-HCl, pH 8.3, 50 mM KCl, 1.5 mM MgCl 2, 0.01% [w / v] gelatin) containing 200 µM of each dNTP, 2.5 units of AmpliTaq polymerase (Perkin-Elmer, Cetus, USA) and 100 pmol of each primer.

The PCR reaction was performed using a DNA thermal cycler (Landgraf, Germany). A incubation at 94 ° C for 1 min followed by thirty PCR cycles using a denaturation cycle profile at 94 ° C for 30 s, pairing at 60 ° C for 1 min and extension at 72 ° C for 2 min. Five aliquots of - \ mul of the amplification product are analyzed by electrophoresis in 0.7% agarose gels (NuSieve, FMC). The appearance of a DNA fragment size of 1.4 kb indicated that amplification of the gene segment was adequate.

Subcloning of PCR fragments

Aliquots of 45- \ mul of PCR products generated as described above were purified using the QIAquick PCR purification kit (Qiagen, USA) according to the manufacturer's instructions. The purified DNA is eluted in 50 µl of 10 mM Tris-HCl, pH 8.5.

5 µg of pMOL944 and 25 µl of the fragment Purified PCR were digested with SacII and NotI, subjected to 0.8% electrophoresis of low temperature agarose gel gelation (SeaPlaque GTG, FMC) and then the fragments relevant were removed from the gels and purified using the QIAquick Gel extraction kit (Qiagen, USA) according to the manufacturer's instructions The isolated DNA PCR fragment is then joined the pMOL944 digested with SacII-NotI and purified. The union was made during overnight at 16 ° C using 0.5 µg of each DNA fragment, 1 U of T4 DNA ligase and T4 ligase buffer (Boehringer Mannheim, Germany).

The ligated mixture was used to transform the PL2306 of competent B. subtilis . The transformed cells were placed on LBPG-10 µg / ml plates of kanamycin. After 18 hours of incubation at 37 ° C colonies were observed on the plates. Different clones were analyzed by plasmid DNA isolation in the night culture broth.

A positive clone of this type was streaked several times on agar plates as previously used and this clone was called MB594. Clone MB594 was grown overnight in TY-10 µg / ml kanamycin at 37 ° C, and the next day 1 ml of cells was used to isolate plasmid from cells using the Qiaprep Spin Plasmid Miniprep Kit No. 27106 according to the recommendations of the manufacturer for plasmid preparations
B. subtilis . This DNA was sequenced and revealed the DNA sequence corresponding to the mature part of the mannanase, that is, positions 94-404 of the SEC. 3 attached. The mature derived protein is shown in the SEC. 4. It seems that the 3 'end of the mannanase encoded by the sequence of SEC. 1 has been changed to the one shown in the SEC. 3 due to the design of the lower primer used in the PCR. The resulting amino acid sequence is shown in the SEC. 4 and it is clear that the C terminal of the SEC. 2 (SHHVREIGVQFSAADNSSGQTALYVDNVTLR) has changed to the C terminal of the SEC. 4 (IIMLGK).

Media

TY (according to Ausubel, F. M. et al. [Eds.] "Current protocols in Molecular Biology". John Wiley and Sons, nineteen ninety five).

LB agar (according to Ausubel, F. M. et al. [Eds.] "Current protocols in Molecular Biology". John Wiley and Sons, nineteen ninety five).

LBPG is LB agar (see above) complemented with 0.5% glucose and 0.05 M potassium phosphate, pH 7.0

BPX medium is described in EP 0 506 780 (WO 09/9129).

Expression, purification and characterization of Bacillus agaradherens mannanase

Clone MB 594 obtained as described previously in Materials and methods it was grown in 25 x 200 ml of BPX medium with 10 µg / ml kanamycin in two diffuser bottles of 500 ml for 5 days at 37 ° C and 31.4 rad / s (300 rpm).

6500 ml of the culture liquid from the bottle of clone MB 594 (lot 9813) and the pH was adjusted to 5.5. Be added 146 ml of cationic agent (C521) and 292 ml of agent anionic (A130) shaking to get flocculation. The material flocculate was separated by centrifugation using a centrifuge Sorval RC 3B at 942.5 rad / s (9000 rpm) for 20 min at 6 ° C. He supernatant was clarified using Whatman GF / D glass filters and C and finally concentrated on a filtron with a cut-off value of 10 kDa

750 ml of this concentrate was adjusted to pH 7.5 with sodium hydroxide. The clear solution was analyzed by anion exchange chromatography with a column 900 ml Q-Sepharose equilibrated with 50 mmol Tris at pH 7.5. The linked mannanase activity was eluted with a gradient of sodium chloride.

The pure enzyme provided a single band in SDS-PAGE with a molecular weight of 38 kDa. amino acid sequence of the enzyme mannanase, that is, the Translated DNA sequence, shown in SEC. 2.

Determination of kinetic constants

Substrate: locust bean gum and analysis of reducing sugars (PHBAH). Sigma Carob Paste (G-0753).

In kinetic determination with different Carob gum concentrations and incubation for 20 min at 40 ° C at pH 10 a value was obtained

Kcat: 467 / s

K m: 0.08 g / l

PM: 38 kDa

pI (isoelectric point): 4.2

The optimum temperature of the mannanase was 60 ° C

The pH activity profile presented an activity maximum between pH 8 and pH 10.

With DSC (differential scanning calorimetry) a temperature of 77 ° C is obtained as a melting point at pH 7.5 in Tris buffer, which indicates that this enzyme is very thermostable.

When 0.2% of AZCL-carob rubber galactomannan as substrate and an incubation according to indications above 40 ° C, the detergent has excellent compatibility with conventional liquid detergents and good compatibility with conventional powder detergents.

Obtaining Bacillus Mannanase Subtilisis 168

The β-mannanase of Bacillus subtilisis was identified and purified as follows: the homology of the genome of Bacillus subtilis was sought with a sequence of known β-mannanase genes from Bacillus sp (Mendoza et al., Biochemica et Biophysica Acta 1243: 552-554, 1995). The coding region of y dhT , whose product was unknown, showed a similarity of 58% with the known Bacillus β-mannanase. The following oligonucleotides were designed to amplify sequence coding for the mature part of the putative β-mannanase: 5'-GCT CAA TTG GCG CAT ACT GTG TCG CCT GTG-3 'and 5'-GAC GGA TCC CGG ATT CAC TCA ACG ATT GGC G-3 '. The total genomic DNA of Bacillus subtilis strain 1A95 was used as a template to amplify the mature ydhT region using the aforementioned primers. PCR is performed with the GENE-AMP PCR kit with AMPLITAQ DNA Polymerase (Perkin Elmer, Applied Biosystems, Foster City, CA). After an initial melting period of 5 min at 95 ° C, 25 cycles of the following program were executed: fusion at 95 ° C for 1 min, mating at 55 ° C for 2 min and extension at 72 ° C for 2 min After the last cycle, the reaction was maintained at 72 ° C for 10 min to complete the extension. The PCR products were purified using the QIAquick PCR purification kit (Qiagen, Chatsworth, CA).

The mature and DHT amplified strain 1A95 Bacillus subtilis region was inserted into the expression vector pPG1524 (previously described) as follows: The 1028bp amplified fragment was digested with Mfe I and Bam H I. The expression vector pPG1527 was digested with Eco R I and Bam H I. Restriction products were purified with the QIAquick PCR purification kit (Qiagen, Chatsworth, CA). The two fragments were ligated using T4 DNA ligase (13 h, 16 ° C) and were used to transform the competent E. coli strain DH5-?. Ampicillin resistant colonies were cultured for DNA preparations. The DNA was then identified by restriction analysis. Plasmid pPG3200 contains the mature region of the gene and dhT . Plasmid pPG3200 was then used to transform competent strain PG 632 of Bacillus subtilis (Saunders et al., 1992).

Seven bacillus subtilis kanamycin-resistant clones and a PG 632 control clone were taken and grown in 20 ml of 20/20/5 medium (20 g / l tryptone, 20 g / l yeast extract, 5 g / l of NaCl) supplemented with 1 ml of 25% maltrin, 120 µl of 10 mM MnCl2 and 20 µl of 50 mg / ml kanamycin. The clones were grown overnight in 250 ml diffuser bottles with stirring at 26.2 rad / s (250 rpm) and 37 ° C to express the protein. The cells were centrifuged at 1466.1 rad / s (14,000 rpm) for 15 minutes. One µl of each supernatant was diluted in 99 µl of 50 mM sodium acetate (pH 6.0). One µl of this dilution was titrated using the beta-Mannazyme endo-1,4-β-mannanase tablets (Megazyme, Ireland) according to the manufacturer's instructions. The absorbance at 590 nm was read on a Beckman DU640 spectrophotometer. Clone 7 has a maximum absorbency of 1.67. The PG632 control has no absorbency at 590 nm.

The supernatant was analyzed by SDS-PAGE on a 10-20% Tris-glycine gel (Novex, San Diego, Ca) to confirm the expected protein size of 38 kDa. Samples were prepared as indicated below. A sample of 500 µl of the clone and dhT 7 and PG 632 supernatants were precipitated with 55.5 µl of 100% trichloroacetic acid (Sigma), washed with 100 µl of 5% trichloroacetic acid, resuspended in 50 µl of buffer for Tris-glycine SDS samples (Novex) and the mixture was boiled for five minutes. One µl of each sample was electrophoresed in the gel at 30 mA for 90 minutes. A large 38 kDa protein band was observed for the clone and dhT 7.

A fermentation of 10 l of clone and dhT 7 of Bacillus subtilis was carried out in a B. Braun Biostat C fermenter. The fermentation conditions were as follows: the cells were cultured for 18 hours in a rich medium similar to 20/20/5 at 37 ° C. At the end of the fermentation cycle the cells were removed and the supernatant was concentrated to 1 liter using a tangential flow filtration system. The final yield of β-mannanase in the concentrated supernatant was 3 g / l.

The purification of the fermentation supernatant β-mannanase It was performed as follows: 500 ml of supernatant was centrifuged at 1047.2 rad / s (10,000 rpm) for 10 min at 4 ° C. He centrifuged supernatant was then dialyzed overnight at 4 ° C in two 4 l loads of 10 mM potassium phosphate (pH 7.2) at through Spectrapor ™ 12,000-14,000 mol. weight with membrane with limiting pore size (spectrum). The supernatant dialysate was centrifuged at 1047.2 rad / s (10,000 rpm) for 10 min at 4 ° C. An anion exchange column of 200 ml Q was equilibrated Sepharose ™ fast flow (Pharmacia) with 1 liter of phosphate 10 mM potassium (pH 7.2) at 20 ° C and 300 ml of supernatant was charged in the column Two pass flow fractions of 210 ml (sample A) and 175 ml (sample B). The two fractions were evaluated as before, unless the samples were diluted with 199 µl of 50 mM sodium acetate (pH 6.0), presenting absorbencies of 0.38 and 0.52, respectively. 2 \ mul added from each sample to 8 µl of sample buffer Tris-glycine SDS (Novex, CA) and remained in boil for 5 min. The resulting samples were subjected to electrophoresis in a Tris-glycine gel at 10-20% (Novex, Ca) at 30 mA for 90 minutes. Be observed a main band corresponding to 38 kDa in each sample which comprised more than 95% of the total protein. A BCA (Pierce) protein determination in both samples according to manufacturer's instructions and using albumin as a standard for bovine serum Samples A and B contained 1.3 mg / ml and 1.6 mg / ml of β-mannanase, respectively. The identity of the protein was confirmed by mass spectrometry type electrospray and amino acid sequence analysis terminals

Purified β-mannanase samples were used to identify enzyme activity as follows: in all determinations, beta-Mannazyme endo-1,4-β-mananase tablets (Megazyme, Ireland) were used as described. previously. The activity in the pH range of 3.0-9.0 was determined with 50 mM citrate phosphate buffer, the activity at pH 9.5 was determined with 50 mM CAPSO buffer (Sigma) and the activity in the pH range of 10.0-11.0 was determined with 50 mM CAPS buffer. The optimum pH for Bacillus subtilis β-mannanase was found to be 6.0-6.5. Activity profiles as a function of temperature were performed in 50 mM citrate phosphate buffer (pH 6.5). The enzyme showed an optimal activity at 40-45 ° C. Bacillus subtilis β-mannanase retained a significant activity below 15 ° C and above 80 ° C. The specific activity against β-1,4-galactomannan was found to be 160,000 µm / min.mg of β-mannanase using beta-Mannazyme endo-1,4-β-mannanase tablets (Megazyme, Ireland) according to The manufacturer's instructions. The nucleotide and amino acid sequences of the β-mannanase from Bacillus subtilisis are shown in the SEC. 5 and 6

Mannanase is incorporated into the compositions of the invention preferably at a level of 0.0001% to 2%, more preferably from 0.0005% to 0.5%, more preferably from 0.001% to 0.1% pure enzyme by weight of the composition.

The enzyme of the invention comprises, in addition to the enzyme core comprising the catalytically active domain, also a cellulose binding domain (CBD), being the domain of cellulose binding and the nucleus (the catalytically active domain) of the enzyme operably linked. The cellulose binding domain (CBD) may exist as an integral part of the encoded enzyme or a CBD from another origin can be introduced into the enzyme to create A hybrid enzyme In this context, the term "binding domain a cellulose "should be understood as defined by Tomme et al. in "Cellulose-Binding Domains: Classification and Properties "in" Enzymatic Degradation of Insoluble Carbohydrates ", John N. Saddler and Michael H. Penner (Eds.), ACS Symposium Series, No. 618, 1996. This definition classifies more than 120 cellulose binding domains distributed in 10 families (I-X) and shows that CBDs are found in different enzymes such as cellulases, xylanases, mannanases, arabinofuranosidases, acetyl esterases or chitinases. CBDs they have also been found in algae, p. eg, in the red seaweed Porphyra purpurea, as a non-hydrolytic polysaccharide binding protein (see Tomme et al., op.cit.) However, most CBDs they come from cellulases and xylanases and are found in N and C Protein terminals or are internal. The hybrid enzymes are known in the art (see, e.g., WO 90/00609 and WO 95/16782) and can be prepared by transformation into a host cell of a DNA structure comprising at least one DNA fragment that encodes the bound cellulose binding domain, with or without a connector, to a DNA sequence that encodes the enzyme mannanase and culturing the host cell to express the fused gene. The hybrid enzymes can be described by the following formula:

CBD - MR - X

where CBD is the region N-terminal or C-terminal of a amino acid sequence corresponding to at least the domain of cellulose binding; MR is the intermediate region (the connector), and can be a joint, or a short bond group preferably of about 2 to about 100 carbon atoms, plus preferably from 2 to 40 carbon atoms; or is preferably of about 2 to about 100 amino acids, more preferably from 2 to 40 amino acids; and X is a region N-terminal or C-terminal enzyme of the invention.

The enzymes mentioned above may have any suitable origin such as, e.g. eg vegetable, animal, Bacterial, fungal or yeast. The origin can also be mesophilic or extremophilic (psychrophilic, psychotropic, thermophilic, barophilic, alkalophilic, acidophilic, halophilic, etc.). Can be used purified or unpurified forms of these enzymes. In the currently it is common practice to modify wild enzymes through protein or genetic engineering techniques to optimize its effectiveness in the cleaning compositions of the invention. By For example, the variants can be designed to increase the Enzyme compatibility with respect to the ingredients of use common in these compositions. Alternatively, the variant is can design in such a way that the optimum pH, the stability of the bleach or chelator, the catalytic activity and the like of the enzyme is suitable for the cleaning application particular.

In particular, special attention should be given to oxidation sensitive amino acids for the stability of bleach and in the case of surface charges for surfactant compatibility. The point can be modified. isoelectric of these enzymes replacing some amino acids loaded so that, p. eg, an increase in the isoelectric point may help improve compatibility with surfactants anionic The stability of enzymes can also be improved creating, p. eg, additional salt bridges to make the Metal binding sites improve chelator stability.

The polymer for dirt release

The laundry detergent composition of The present invention generally comprises from 0.0001% to 20%, preferably from 0.001% to 15%, more preferably from 0.01% to 10%, by weight of a polymer for the release of dirt for Polyethyleneimine type cotton fabrics. The polymers for dirt release for cotton type fabrics Preferred polyethyleneimines are agents to release dirt of water-soluble modified polyamine cotton fabrics or dispersibles comprising a polyamine main chain corresponding to the formula described in WO97 / 42288, presented on April 25, 1997 by Procter & Gamble:

5

which has a polyamine formula modified V (n + 1)} W_ {m} Y_ {n} Z or a main chain polyamine of formula:

6

which has a polyamine formula modified V (n-k + 1)} W_m Y_ {n} Y 'k Z, in where k is less than or equal to n, said polyamine main chain before modification it has a molecular weight greater than approximately 200 daltons, in where

i)

 the V units are terminal units of formula:

7

ii)

the W units are main chain units of formula:

8

iii)

the Y units are branching units of formula:

9

iv)

the Z units are terminal units of formula:

10

where the union units R of the main chain are selected from the group consisting of C 2 -C 12 alkylene, alkenylene C 4 -C 12, hydroxyalkylene C 3 -C 12, dihydroxyalkylene C 4 -C 12, dialkylarylene C 8 -C 12, - (R 1 O) x R 1 -,  - (R 1 O) x R 5 (OR 1) x -, - (CH 2 CH (OR 2) CH 2 O) z - (R 1 O) and R 1 (OCH 2 CH (OR 2) CH 2) w -,  -C (O) (R 4) r C (O) -, -CH 2 CH (OR 2) CH 2 -, and mixtures of the themselves; wherein R 1 is alkylene C 2 -C 6 and mixtures thereof; R2 is hydrogen, - (R 1 O) x B, and mixtures thereof; R 3 is C 1 -C 18 alkyl, arylalkyl C 7 -C 12, alkyl substituted aryl C 7 -C 12, aryl C 6 -C 12, and mixtures thereof; R 4 it is C 1 -C 12 alkylene, alkenylene C 4 -C 12, arylalkylene C 8 -C 12, arylene C 6 -C 10, and mixtures thereof; R 5 is C 1 -C 12 alkylene, hydroxyalkylene C 3 -C 12, dihydroxyalkylene C 4 -C 12, dialkylarylene C_ {8} -C_ {12}, -C (O) -, -C (O) NHR 6 NHC (O) -, -R 1 (OR 1) -, -C (O) (R 4) r C (O) -, -CH 2 CH (OH) CH 2 -, -CH 2 CH (OH) CH 2 O (R 1 O) and R 1 -OCH 2 CH (OH) CH 2 -, and mixtures thereof; R 6 is alkylene C 2 -C 12 or arylene C 6 -C 12; E units are selected from group consisting of hydrogen, C 1 -C 22 alkyl, C 3 -C 22 alkenyl, arylalkyl C 7 -C 22, hydroxyalkyl C 2 -C 22, - (CH 2) p CO 2 M, - (CH 2) q SO 3 M, -CH (CH 2 CO 2 M) CO 2 M, - (CH 2) p PO 3 M, - (R 1 O) x B, -C (O) R3, and mixtures thereof; as long as when any unit E of a nitrogen is a hydrogen, said Nitrogen is also not an N-oxide; B is hydrogen, C 1 alkyl -C 6, - (CH 2) q SO 3 M, - (CH 2) p CO 2 M, - (CH 2) q (CHSO 3 M) CH 2 SO 3 M, - (CH 2) q (CHSO 2 M) -CH 2 SO 3 M, - (CH 2) p PO 3 M, -PO 3 M, and mixtures of the themselves; M is hydrogen or a water-soluble cation in quantity enough to satisfy the load balance; X is an anion water soluble;

k and k 'have the value of 1 to approximately fifteen; m has the value of 4 to about 400; n has the value of 0 to about 200; p has the value from 1 to 6, which has the value from 0 to 6; r has the value 0 or 1; w has the value 0 or 1; x has the value from 1 to 100; and has the value from 0 to 100; z has the value 0 or one.

These polyamines comprise main chains which can be linear or cyclic. The main chains polyamines can also comprise to a greater or lesser degree branching chains In general, the main chains polyamines described herein are modified from such that each nitrogen in the polyamine chain is described at then as a substituted, quaternized, oxidized or combinations thereof.

For the purposes of the present invention the term "modification" is defined as the replacement of a hydrogen atom of the unit -NH of the main chain by a unit E (substitution), the quaternization of a nitrogen from the main chain (quaternized) or the oxidation of a nitrogen from the main chain to form N-oxide (oxidized). The terms "modification" and "substitution" are used in a way interchangeable when referring to the process of replacing a hydrogen atom attached to a main chain nitrogen by a unit E. Quaternization or oxidation can take place in some cases without substitution, but preferably substitution It is accompanied by the oxidation or quaternization of at least one main chain nitrogen.

The main linear polyamine chains or non-cyclic comprised by agents to release dirt in Cotton fabrics of the present invention have the formula general:

eleven

said main chains before be modified include primary, secondary amine nitrogens and tertiary joined by R units "union". The chains major cyclic polyamines comprising the agents for release dirt on cotton fabrics of the present invention they have the formula general:

12

said main chains before be modified include primary, secondary amine nitrogens and tertiary joined by units R "of Union".

In the present invention, the nitrogens of the primary amine comprising the main chain or the chain of branching once modified are defined as units "terminals" V or Z. For example, when an amine residue primary located at the end of the polyamine main chain or in the branching chain that has the structure

[H 2 N-R] -

is modified according to the present invention, hereafter defined as a unit V "terminal" or simply as a V unit. However, in the present invention, some or all primary amine residues may remain unmodified subject to the restrictions that are will be described later herein. Because of his position in the main chain, these primary amine residues do not modified are still "terminal" units. So similar, when a primary amine residue located at the end of the polyamine main chain that has the structure

-NH_ {2}

is modified according to the present invention, hereafter defined as a unit Z "terminal" or simply as a Z unit. This unit can remain unchanged subject to the restrictions that are will be described later herein memory.

Similarly, the amine nitrogens secondary comprising the main chain or the chain of branching once modified are defined as W units of main string ". For example, when you modify a remainder of secondary amine, the main constituent of the chains main and branching chains of the present invention that has the structure

13

according to the present invention, as far as successive is defined as a "main chain" W unit or simply as a W unit. However, in the present invention some or all of the secondary amine residues may remain without Modify. Due to their position in the "main chain", these secondary amine residues remain "chain" units principal".

Similarly, once modified the tertiary amine nitrogens comprising the main chain or the branching chain, hereinafter referred to as "branching" units. For example, when a rest of tertiary amine, which is the branching point of the chain main polyamine or other branching chains or rings that have the structure

14

is modified according to the present invention, hereinafter defined as unit Y of "branching" or simply as a Y unit. However, in the present invention some or all tertiary amine residues They can remain unmodified. Due to its position in the chain main, these unmodified tertiary amine residues follow being "branching" units. The R units associated with the nitrogen of the unit V, W and Y that serve to unite the polyamine nitrogens are described later in the Present memory.

The final modified structure of polyamines of the present invention can therefore be represented by the general formula

V (n + 1)} W_ {m} Y_ {n} Z

for polyamine polymers for the release of dirt in linear cotton fabrics and by the formula general

V (n-k + 1)} W_ {m} Y_ {n} Y '{} k

for polyamine polymers cyclic for the release of dirt in cotton fabrics. In the case of polyamines comprising rings, a Y 'unit of formula

fifteen

serves as a branching point of a main chain or branching ring. For each unit Y 'there is a unit Y that has the formula

16

which will form the junction point of ring with the polymeric main chain or branching. Only if the main chain is a ring complete, the polyamine main chain has the formula

17

and therefore does not understand any terminal unit Z and has the formula

V_ {n-k} W_ {m} Y_ {n} Y '{} _ {k}

where k is the number of units of branching that form the ring. Preferably the chains Main polyamines of the present invention do not comprise rings

In the case of non-cyclic polyamines, the quotient between index n and index m indicates the relative degree of branch. A fully linear modified polyamine does not branched according to the present invention has the formula

VW_ {m} Z

that is, n is equal to 0. How much the higher the value of n (the lower the m: n ratio), the higher the degree of branching in the molecule. Typically the value of m is in the range of at least 4 to about 400, although values greater than m are also preferred, especially when the index n value is very low or almost 0.

Each polyamine nitrogen, regardless of whether it is primary, secondary or tertiary, a once modified according to the present invention, it is further defined as belonging to a member of one of the three general classes: with simple, quaternized or oxidized replacement. Nitrogen units of unmodified polyamine are classified into units V, W, Y or Z depending on whether they are primary, secondary or Tertiary That is, for the purposes of the present invention the Unmodified primary amine nitrogens are units V or Z, the unmodified secondary amine nitrogens are W units and the Unmodified tertiary amine nitrogens are Y units.

The modified primary amine residues are defined as "terminal" units V that have one of the three shapes:

to)

units with simple replacement that have the structure:

18

b)

quaternized units that have the structure:

         \ vskip1.000000 \ baselineskip
      

19

where X is a proper counterion that provides load balancing Y

C)

oxidized units that have the structure:

         \ vskip1.000000 \ baselineskip
      

twenty

The modified secondary amine residues are defined as "main chain" W units having one of The following three ways:

to)

units with simple replacement that have the structure:

         \ vskip1.000000 \ baselineskip
      

twenty-one

b)

quaternized units that have the structure:

         \ vskip1.000000 \ baselineskip
      

22

where X is a proper counterion that provides load balancing Y

C)

oxidized units that have the structure:

         \ vskip1.000000 \ baselineskip
      

2. 3

The modified tertiary amine residues are defined as "branching" Y units that have one of the Three following ways:

to)

Unmodified units that have the structure:

         \ vskip1.000000 \ baselineskip
      

24

b)

quaternized units that have the structure:

25

where X is a proper counterion that provides load balancing Y

C)

oxidized units that have the structure:

26

Certain modified primary amine residues are defined as "terminal" Z units that have one of the three following forms:

to)

units with simple replacement that have the structure:

27

b)

quaternized units that have the structure:

28

where X is a proper counterion that provides load balancing Y

C)

oxidized units that have the structure:

29

When any position in a nitrogen does not is substituted or modified, it is understood that hydrogen will be replaced by E. Thus, for example, a primary amine unit that comprises an unit E in the form of a hydroxyethyl moiety is a terminal unit V having the formula (HOCH 2 CH 2) HN-.

In the present invention there are two types of chain end units, units V and units Z. The "terminal" Z unit is derived from a primary amine moiety terminal of the structure -NH_ {2}. The main chains non-cyclic polyamines according to the present invention comprise only one Z unit while cyclic polyamines can not understanding Z units. The "terminal" Z unit can be replaced by any of the units E described below herein, except when the Z unit is modified to form an N-oxide. If the nitrogen in unit Z is oxidized to N-oxide, the nitrogen must be modified and, therefore, E cannot Be a hydrogen.

The polyamines of the present invention they comprise "binding" R units of the main chain that they serve to unite the nitrogen atoms of the main chain. The R units comprise units that in the present invention receive the name of R units "hydrocarbyl" and R units "oxy". The "hydrocarbyl" R units are alkylene C 2 -C 12, alkenylene C 4 -C 12, C 3 -C 12 hydroxyalkylene where the hydroxyl moiety can have any position in the chain of R units except carbon atoms directly bound to the nitrogens of the polyamine main chain; C 4 -C 12 dihydroxyalkylene where the hydroxyl moieties can occupy either of the two atoms of carbon from the chain of R units except carbon atoms directly bound to the main chain nitrogen polyamine; C 8 -C 12 dialkylarylene which for the purposes of the present invention are arylene moieties that they have two alkyl substituent groups as part of the chain of Union. For example, a dialkylarylene unit has the formula

30

although the unit does not need to be 1,4-substituted, but it can also be 1,2 or 1,3 C2-C12 substituted alkylene, preferably ethylene, 1,2-propylene, and mixtures of the same, more preferably ethylene. R units "oxy" understand - (R 1 O) x R 5 (OR 1) x -, -CH 2 CH (OR 2) CH 2 O) z (R 1 O) and R 1 (OCH 2 CH (OR 2) 2) CH 2) w -,  -CH 2 CH (OR 2) CH 2 -, - (R 1 O) x R 1 -, and mixtures thereof. The Preferred R units are alkylene C 2 -C 12, hydroxyalkylene C 3 -C 12, dihydroxyalkylene C 4 -C 12, dialkylarylene C 8 -C 12, - (R 1 O) x R 1 -, -CH 2 CH (OR 2) CH 2 -, - (CH 2 CH (OH) CH 2 O) z (R 1 O) and R 1 (OCH 2 CH- (OH) CH 2 }) w -,  - (R 1 O) x R 5 (OR 1) x -, most preferred R units are alkylene C 2 -C 12, hydroxyalkylene C 3 -C 12, dihydroxyalkylene C 4 -C 12, - (R 1 O) x R 1 -,  - (R 1 O) x R 5 (OR 1) x -, - (CH 2 CH (OH) CH 2 O) z (R 1 O) and R 1 (OCH 2 CH- (OH) CH 2 }) w -,  and mixtures thereof, even more preferred R units are C 2 -C 12 alkylene, hydroxyalkylene C 3, and mixtures thereof, being the most preferred C 2 -C 6 alkylene. The main chains Most preferred of the present invention comprise at least 50% of R units that are ethylene.

The R1 units are alkylene C 2 -C 6, and mixtures thereof, preferably ethylene.

R2 is hydrogen or - (R 1 O) x B, preferably hydrogen.

R 3 is alkyl C 1 -C 18, arylalkylene C 7 -C 12, alkyl substituted aryl C 7 -C 12, aryl C 6 -C 12, and mixtures thereof, preferably C 1 -C 12 alkyl, C 7 -C 12 arylalkylene, more preferably C 1 -C 12 alkyl and most preferably methyl. The R3 units serve as part of the E units described later herein.

R 4 is alkylene C_ {1} -C_ {12}, alkenylene C 4 -C 12, arylalkylene C 8 -C 12, arylene C 6 -C 10, preferably alkylene C 1 -C 10, arylalkylene C 8 -C 12, more preferably alkylene C 2 -C 8 and most preferably ethylene or butylene

R 5 is C 1 -C 12 alkylene, C 3 -C 12 hydroxyalkylene, C 4 -C 12 dihydroxyalkylene, C 8 -C 12 dialkylarylene, -C (O) -, -C (O) NHR 6 NHC (O) -, -C (O) (R 4) r C (O) -, -R 1 (OR 1) -, -CH 2 CH (OH) CH 2 O (R 1 O) and R 1 OCH 2 CH (OH) CH_ {2 -, -C (O) (R 4) r C
(O) -, -CH 2 CH (OH) CH 2 -, R 5 is preferably ethylene, -C (O) -, -C (O) NHR 6 NHC (O) -, -R 1 (OR 1) -, -CH 2 CH (OH) CH 2 -, -CH 2 CH (OH) CH 2 O (R 2 1 O) and R 1 OCH 2 CH- (OH) CH 2 -, more preferably -CH 2 CH (OH) CH 2 -.

R 6 is alkylene C 2 -C 12 or arylene C_ {6} -C_ {12}.

Preferred "oxy" R units are also defined in terms of the units R1, R2 and R5. The Preferred "oxy" R units comprise R1 units, Preferred R2 and R5. Agents to release dirt in preferred cotton fabrics in the present invention comprise at least 50% of R1 units that are ethylene. The units Preferred R 1, R 2 and R 5 are combined with R units "oxy" to obtain the preferred "oxy" R units of the Following way:

i)

 The replacement of the most preferred R5 units in - (CH 2 CH 2 O) x R 5 (OCH 2 CH 2) x - provides - (CH 2 CH 2 O) x CH 2 CHOHCH 2 (OCH 2 CH 2) x -.

ii)

The replacement of the preferred R1 and R2 units in - (CH 2 CH (OR 2) CH 2 O) z - (R 1 O) and R 1 O (CH 2 CH ( OR 2) CH 2) w - provides - (CH 2 CH (OH) CH 2 O) z - (CH 2 CH 2 O) and CH 2 CH 2 O (CH 2) CH (OH) CH 2) w -.

iii)

The replacement of the preferred R2 units in -CH 2 CH (OR 2) CH 2 - provides

-CH 2 CH (OH) CH 2 -.

The units E are selected from the group consisting of hydrogen, C 1 -C 22 alkyl, C 3 -C 22 alkenyl, C 7 -C 22 arylalkyl, C 2 hydroxyalkyl -C 22, - (CH 2) p CO 2 M, - (CH 2) q SO 3 M, -CH (CH 2 CO 2 M) CO 2 M, - (CH 2) p PO 3 M, - (R 1 O) m
B, -C (O) R 3, preferably hydrogen, C 2 -C 22 hydroxyalkylene, benzyl, C 1 -C 22 alkylene, - (R 1 O) {m} B, -C (O) R3,
- (CH 2) p CO 2 M, - (CH 2) q SO 3 M, -CH (CH 2 CO 2 M) CO 2 M, more preferably C 1 -C 22 alkylene, - (R 1 O) x B, -C (O) R 3, - (CH 2) _ {p} CO 2 M, - (CH 2) q SO 3 M, -CH (CH 2 CO 2 M) CO 2 M, most preferably C_ alkylene {1 -C 22, - (R 1 O) x B, and -C (O) R 3. If no modification or substitution has been made in a nitrogen, then the hydrogen atom remains the remainder representing E.

         \ newpage
      

The E units do not comprise an atom of hydrogen when the units V, W or Z are oxidized, that is, the Nitrogens are N-oxides. For example, the main chain or branching chains do not comprise units of the following structure:

31

In addition, the units E do not include remains carbonyl attached directly to a nitrogen atom when V, W or Z units are oxidized, that is, nitrogen is N-oxides According to the present invention, the unit E -C (O) R3 is not bound to a nitrogen modified to N-oxide, that is, there are no N-oxide amides with the structure

32

or combinations of same.

B is hydrogen, C 1 -C 6 alkyl, - (CH 2) q SO 3 M, - (CH 2) p CO 2 M, - (CH 2) q - (CHSO 3 M) CH 2 SO 3 M, - (CH 2) q (CHSO 2 M) CH 2 SO 3 M, - (CH 2) p PO 3 M, -PO 3 M, preferably hydrogen, - (CH 2) q SO 3 M, - (CH 2) q (CHSO 3 M) CH 2 SO 3 M,
- (CH 2) q - (CHSO 2 M) CH 2 SO 3 M, more preferably hydrogen or - (CH 2) q SO 3 M.

M is hydrogen or a water-soluble cation in sufficient quantity to satisfy the load balance. For example, a sodium cation also satisfies - (CH 2) p CO 2 M, and - (CH 2) q SO 3 M, providing moieties - (CH 2 }) pCO2 Na and
- (CH 2) q SO 3 Na. More than one monovalent cation, (sodium, potassium, etc.) can be combined to meet the required chemical load balance. However, more than one anionic group may exhibit charge equilibrium by a divalent cation or more than one monovalent cation may be necessary to meet the loading requirements of a polyanionic radical. For example, a moiety - (CH 2) p PO 3 M substituted with sodium atoms has the formula
- (CH 2) p PO 3 Na 3. Divalent cations such as calcium (Ca 2+) or magnesium (Mg 2+) can be substituted or combined with other suitable monovalent water-soluble cations. The preferred cations are sodium and potassium and the most preferred is sodium.

X is a water-soluble anion such as chlorine (Cl -), bromine (Br -) or iodine

(I -) or X can be any radical with negative charge such as sulfate (SO4 2-) and methosulfate (CH 3 SO 3 -).

The indices of the formula have the following values: p has the value from 1 to 6, q has the value from 0 to 6; r it has the value 0 or 1; w has the value 0 or 1, x has the value of 1 to 100; and has the value from 0 to 100; z has the value 0 or 1; what is less than or equal to the value of n; m has the value of 4 a about 400, n has the value of 0 to about 200; m + n has the value of at least 5.

Agents to release tissue dirt Preferred cotton in the present invention comprise chains main polyamines where less than about 50% of R groups comprise "oxy" R units, preferably less of about 20%, more preferably less than 5% and with maximum Preferably, the R units do not comprise "oxy" R units.

Agents to release tissue dirt of cotton that do not include more preferred R "oxy" units they comprise polyamine main chains where less than 50% of R groups comprise more than 3 carbon atoms. For example, ethylene, 1,2-propylene and 1,3-propylene comprise 3 or less carbon atoms and are the preferred "hydrocarbyl" R units. That is to say, when the R units of the main chain are alkylene C 2 -C 12, alkylene is preferred C 2 -C 3, with the most preferred being ethylene.

Agents to release tissue dirt of cotton of the present invention comprise main chains Homogeneous and non-homogeneous modified polyamines, where 100% or less of the -NH units are modified. At the moment invention the term "homogeneous polyamine main chain" It is defined as a polyamine main chain that has units R which are the same (eg, all ethylene). However, the definition of uniformity does not exclude polyamines comprising other foreign units comprising the polymeric main chain and that are present due to an impurity of the synthesis method chosen chemistry. For example, those skilled in the art know that as an "initiator" of the synthesis of polyethyleneimines, it is possible to use ethanolamine, so a sample of polyethyleneimine that comprises a hydroxyethyl moiety resulting from the "initiator" of the polymerization would be considered to comprise a main chain homogeneous polyamine of the present invention. A main chain polyamine comprising all R ethylene units in which no there are branching units Y is a main chain homogeneous A polyamine main chain comprising all the R ethylene units is a homogeneous main chain regardless of the degree of branching or the number of cyclic ramifications present.

In the present invention the term "chain non-homogeneous polymeric principal "refers to chains main polyamines that are a mixture of R units of different lengths and R units of different types. For example, a non-homogeneous main chain comprises R units that are a mixture of ethylene units and units 1,2-propylene. In the present invention it is not necessary a mixture of R units "hydrocarbyl" and R units "oxy" to obtain a non-homogeneous main chain. The proper handling of these "unit chain lengths R "allows the formulator to modify the solubility and efficacy of the agents to release dirt on cotton fabrics from the present invention

Polymers for dirt release in preferred cotton fabrics in the present invention comprise homogeneous polyamine main chains that are total or partially substituted by polyethylene oxides, total amines or partially quaternized, totally or partially oxidized nitrogen to N-oxide, and mixtures thereof. However, not all Nitrogen from the main chain amine must be modified in the same way, leaving the choice of modification to the specific needs of the formulator. The degree of ethoxylation is also determined by the specific requirements of the formulator.

Preferred polyamines comprising the Main chain of the compounds of the present invention are generally polyalkyleneamines (PAAs), polyalkyleneimines (PAIs), preferably polyethyleneamines (PEAs), polyethyleneimines (PEIs), or PEAs or PEIs joined by residues that have R units longer than the PAAs, PAIs, PEAs or precursor PEIs. A polyalkyleneamine (PAA) Common is tetrabutylenpentamine. PEAs are obtained through reactions involving ammonia and ethylene dichloride, followed by a fractional distillation. The common PEAs obtained are triethylenetetramine (TETA) and tetraethylenepentamine (TEPA). Above of pentamines, that is, hexamines, heptamines, octamines and, possibly nonamines, the cogenically derived mixture not seems to be separated by distillation and may include other materials such as cyclic amines, particularly piperazines. Too cyclic amines with side chains may be present with nitrogen atoms See US Patent 2,792,372, granted to Dickinson on May 14, 1957, which describes the PEAs preparation.

The main chains of amino polymer Preferred comprise R units that are alkylene units (ethylene) C2, also known as polyethyleneimines (PEIs). Preferred PEIs have at least moderate branching, it is say the ratio m: n is less than 4: 1, although the PEIs that have an m: n ratio of about 2: 1 are more preferred. The preferred main chains, before modification have the General Formula:

33

where m and n are equal according to You have previously defined herein. PEIs preferred, before modification, will have a molecular weight greater than about 200 daltons.

The relative proportions of amine units primary, secondary and tertiary in the polyamine main chain, especially in the case of IEPs, they will vary depending on the form of preparation Each hydrogen atom attached to each of nitrogen of the polyamine main chain represents a possible site for subsequent replacement, quaternization or oxidation.

These polyamines can be prepared, by example, by polymerization of ethyleneimine in the presence of a catalyst such as carbon dioxide, sodium bisulfite, acid sulfuric acid, hydrogen peroxide, hydrochloric acid, acetic acid, etc. Specific methods to prepare these main chains polyamines are described in the patents US 2,182,306, granted to Ulrich et al. on 5 of December 1939; US-3,033,746, granted to Mayle and cabbage. on May 8, 1962; US 2,208,095, granted to Esselmann et al. on July 16, 1940; US 2,806,839, issued to Crowther on 17 September 1957; and US 2,553,696, granted to Wilson on May 21, 1951.

Examples of polymers for the release of the dirt on modified cotton fabrics of the present invention which comprise PEIs are those illustrated in the formulas I-V:

Formula I illustrates a polymer for dirt release in preferred cotton fabrics that comprises a main PEI chain where all the nitrogen replaceable are modified by replacing hydrogen with a polyoxyalkyleneoxy unit, - (CH2CH2O) 20H, which It has the formula:

3. 4

Formula II illustrates a polymer for release of dirt on cotton fabrics comprising a PEI main chain where all replaceable nitrogen are modified by replacing hydrogen with a unit polyoxyalkyleneoxy, - (CH 2 CH 2 O) 7 H, which has the formula

35

This is an example of a polymer for dirt release on cotton fabrics that is totally modified by a type of rest.

Formula III illustrates a polymer for release of dirt on cotton fabrics comprising a PEI main chain where all primary amine nitrogens replaceable are modified by replacing hydrogen with a polyoxyalkyleneoxy unit, - (CH 2 CH 2 O) 7 H and the molecule is then modified by a subsequent oxidation of all primary and secondary oxides oxidizable to N-oxide, said agent having to release dirt from cotton fabrics the formula

36

Formula IV illustrates a polymer for the release of dirt in cotton fabrics comprising a PEI main chain where all the hydrogen atoms of the main chain are substituted and some amine units of the main chain are quaternized. The substituents are polyoxyalkyleneoxy units, - (CH 2 CH 2 O) 7
H, or methyl groups. The polymer for the release of dirt in cotton fabrics with modified PEI has the formula

37

Formula V illustrates a polymer for release of dirt on cotton fabrics comprising a PEI main chain in which the main chain nitrogens are modified by substitution (e.g., by - (CH 2 CH 2 O) 7 H or methyl), quaternized, oxidized to N-oxides or combinations thereof. The polymer for the release of dirt in resulting cotton fabrics has the formula

38

In the examples above, not all Nitrogens of a unit class comprise the same modification. The present invention allows the formulator to have a portion of the secondary amine nitrogens ethoxylated and have other nitrogen from secondary amine oxidized to N-oxide. This also applies to the primary amine nitrogen, where the formulator can choose to modify all or part of the primary amine nitrogens with one or more substituents before oxidation or quaternization Any possible combination of E groups in primary and secondary amine nitrogens, except for the restrictions described above herein memory.

The formulator can take advantage of the possibility of modify the polyamine main chains of this invention so that only minimal oxidation of the main chains of the substrate. For example, a "mild" bleaching before or after formulation. For the purposes of the present invention, the expression "tempered of bleaching "is defined as treating the modified polyamine with enough bleaching agent to oxidize the main chain against the formulation conditions. By way of illustration, a polyamine main chain does not necessarily require a total modification by quaternization or N-oxidation to be stable to bleach. When a string sample modified polyamine principal is exposed to a system suitable bleach (for example, sulphonate / perborate nonanoyloxybenzene), any nitrogen in the main chain oxidizable will oxidize under these conditions. However, due to the exact structural properties of the main chain, some or all nitrogens in the pre-whitening treatment may not be seen affected. Once this tempering is done, the formulator can combine the modified polyamine with the bleaching system and be sure that polyamine will not consume most of the agent bleach.

The expert in the art of formulations bleachers will recognize that bleaching tempering has its limitations and that bleaching tempering should not be used anymore weak instead of bleaching the formulation.

In another way, the formulator may wish to add an excess of bleaching agent to the laundry detergent composition during the formulation to carry out a suitable "warm" bleaching in situ during storage and handling of the formulation.

A preferred embodiment of the present The invention involves the use of polyhydroxyamide type surfactant of fatty acid together with the modified polyamines described in the present memory This combination of non-ionic surfactant and modified polyamine is especially useful in low formulations pH, that is, a pH below about 10. The fatty acid polyhydroxyamide suitable for use in Low pH embodiments of the present invention may be combined with other suitable detersive surfactants such as anionic, ampholytic, hybrid ion surfactants, and mixtures of the same.

For the purpose of the present invention, prefer polymer for tissue release of polyethyleneimine type cotton selected from polyethyleneimine 1800E7 and its derivatives of amine oxide, polyethyleneimine 1200E7 and its oxidized and / or quaternized derivatives, polyethyleneimine 600E20, and / or mixtures thereof as described in the Examples 1-4 of WO97 / 42288.

Detergent components

Laundry detergent compositions of the invention must contain at least one detergent component additional. The precise nature of these additional components and their levels of incorporation will depend on the way physics of the composition and nature of the operation of cleaning for which they will be used.

Laundry detergent compositions of the present invention preferably also comprise another detergent ingredient selected from a reinforcing additive of the detergency, especially a zeolite, a sodium tripolyphosphate and / or laminar silicate, a surfactant, preferably a surfactant non-ionic such as alkylethoxylate or alkylmethyl glucamide, a polymer for the release of conventional dirt and / or mixtures of same.

Laundry detergent compositions according to the invention they can be in the form of liquid, paste, gel, bar, tablet, spray, foam, powder or granulate. The granulated compositions may also be in shape "compact" and liquid compositions can also be in "concentrated" form.

The compositions of the invention may be formulated, for example, as detergent compositions for laundry by hand or machine including compositions with laundry additive and compositions suitable for use in the soaking and / or pretreatment of dirty tissues and in the fabric softener compositions added during the cleared up.

When the compositions of the invention are formulated as compositions suitable for use in a method for washing in a washing machine, preferably they contain a surfactant and a detergent builder additive and also one or more detergent components preferably selected from compounds organic polymers, bleaching agents, additional enzymes, defoamers, dispersant, lime soap dispersant, stain suspensions and anti-redeposition agents and inhibitors of corrosion The washing compositions may also contain softeners as supplemental detergent components. These compositions containing a mannanase and a polymer for dirt release in cotton fabrics can provide Advantages of tissue cleaning, stain removal, whiteness and color appearance when formulated as laundry detergent compositions.

The compositions of the invention can also be used as detergent additives in solid or liquid form. These additives are intended to complement or improve the performance of conventional detergent compositions and can be added at any stage of the cleaning process.

If necessary, the density of laundry detergent compositions herein invention may range from 400 to 1200 g / liter, preferably from 500 to 950 g / liter, of the compositions measured at 20 ° C.

The "compact" form of the compositions of the present invention is optimally reflected by the density and, in terms of composition, by the amount of salt of inorganic load Inorganic filler salts are ingredients conventional detergent powder compositions; in the conventional detergent compositions the loading salt is present in significant quantities, typically 17% to 35% by weight of the total composition.

In compact compositions, the loading salt It is present in amounts that do not exceed 15% of the composition total, preferably not exceeding 10% and with maximum Preference not exceeding 5%, by weight of the composition. The inorganic filler salts, such as those used herein compositions, are selected from alkali metal salts and alkaline earth sulfates and chlorides. A preferred loading salt It is sodium sulfate.

Liquid detergent compositions according to the The present invention may also be in "concentrated form", in which case the detergent liquid compositions according to the The present invention will contain a lower amount of water than conventional liquid detergents. Typically the water content of the concentrated liquid detergent is preferably less than 40%, more preferably less than 30% and most preferably less than 20%, by weight of the composition Detergent.

Suitable detergent compounds for use in The present invention is selected from the group consisting of Compounds listed below.

Surfactant system

Preferably, the detergent compositions for washing clothes according to the present invention can also comprise a surfactant system where the surfactant can be selected from nonionic and / or anionic surfactants and / or cationic and / or ampholytic and / or hybrid and / or semipolar ion. Especially, laundry detergent compositions of the The present invention will comprise, in addition to the enzyme mannanase and the polymer for the release of dirt in cotton fabrics, a nonionic surfactant, preferably alkylethoxylated with a chain length from C8 to C20, preferably from C12 to C16, and a degree of ethoxylation from 2 to 9, preferably from 3 to 7, or a alkylmethyl glucamine type surfactant with a length of C8 to C20 alkyl chain, preferably C12 to C18. It has been surprisingly observed that these compositions provide better cleaning capacity, especially cosmetic stains and food, and greater advantages of dirt release.

The other surfactant is typically present at a level of 0.1% to 60% by weight. The levels of more preferred incorporation are from 1% to 35% by weight, with maximum preference of 1% to 30% by weight of the detergent compositions of laundry washing according to the invention.

The surfactant is preferably formulated to that is compatible with the enzymatic components present in the composition. In liquid or gel compositions, the surfactant is formulates most preferably so that it improves, or at least not reduce the stability of the enzymes present in these compositions

The condensates of poly (ethylene oxide), poly (propylene oxide) and poly (butylene oxide) of the alkyl Phenols are suitable for use as the non-ionic surfactant of the surfactant systems of the present invention, the Preferred poly (ethylene oxide) condensates. These Compounds include condensation products of alkylphenols with an alkyl group containing from about 6 to about 14 carbon atoms, preferably of about 8 to about 14 carbon atoms, in linear chain or branched chain configuration with oxide alkylene In a preferred embodiment, the ethylene oxide is present in an equivalent amount of approximately 2 to about 25 moles, more preferably about 3 to approximately 15 moles of ethylene oxide per mole of alkylphenol Commercial non-ionic surfactants of this type include Igepal ™ CO-630, marketed by GAF Corporation; and Triton ™ X-45, X-114, X-100 and X-102, all marketed by Rohm & Haas Company. These surfactants are called generically alkyl phenol alkoxylates (for example alkylphenol ethoxylates).

The condensation products of alcohols primary and secondary aliphatics that are approximately 1 to approximately 25 moles of ethylene oxide are suitable for its use as the non-ionic surfactant of surfactant systems nonionic of the present invention. The alkyl chain of alcohol aliphatic can be linear or branched, primary or secondary and, Generally, it contains about 8 to about 22 carbon atoms Preference is given to condensation products of alcohols with an alkyl group containing about 8 at about 20 carbon atoms, more preferably of about 10 to about 18 carbon atoms, with of about 2 to about 10 moles of ethylene oxide per mole of alcohol They are present in these condensation products from about 2 to about 7 moles of ethylene oxide and most preferably 2 to 5 moles of ethylene oxide per mole of alcohol. Examples of commercial nonionic surfactants of this type include Tergitol ™ 15-S-9 (the alcohol condensation product C 11 -C 15 linear with 9 moles of oxide ethylene), Tergitol ™ 24-L-6 NMW (el alcohol condensation product C 12 -C 14 primary with 6 moles of oxide ethylene with a narrow molecular weight distribution), both marketed by Union Carbide Corporation; Neodol ™ 45-9 (the condensation product of alcohol C 14 -C 15 linear with 9 moles of oxide ethylene), Neodol ™ 23-3 (the condensation product of the linear C 12 -C 13 alcohol with 3.0 moles of ethylene oxide), Neodol ™ 45-7 (the product of condensation of linear C 14 -C 15 alcohol with 7 moles of ethylene oxide), Neodol ™ 45-5 (the alcohol condensation product C 14 -C 15 linear with 5 moles of oxide ethylene) marketed by Shell Chemical Company, Kyro ™ EOB (the alcohol condensation product C 13 -C 15 with 9 moles of ethylene oxide), marketed by The Procter & Gamble Company, and Genapol LA O3O or O5O (the condensation product of alcohol C 12 -C 14 with 3 or 5 moles of oxide ethylene) marketed by Hoechst. The preferred HLB interval in these products it is 8-11 and with maximum 8-10 preference.

         \ newpage
      

Also useful as the nonionic surfactant of The surfactant systems of the present invention are the alkylpolysaccharides described in the patent US 4,565,647, granted to Llenado on January 21, 1986, which have a hydrophobic group containing approximately 6 to about 30 carbon atoms, preferably of about 10 to about 16 carbon atoms, and a polysaccharide, p. eg, a hydrophilic polyglycoside group that contains from about 1.3 to about 10, preferably from about 1.3 to about 3 and with highest preference of about 1.3 to about 2.7, saccharide units. Any saccharide reducer can be used that contains 5 or 6 carbon atoms, such as residues of glucose, galactose and galactosyl can be substituted by the moieties glycosyl (optionally the hydrophobic group is linked to the positions 2, 3, 4, etc. forming, therefore, a glucose or galactose instead of a glycoside or galactoside). The links between saccharides may be, for example, between position one of the additional saccharide units and positions 2, 3, 4 and / or 6 in the preceding saccharide units.

Preferred alkyl polyglycosides have the formula

R 2 O (C n H 2 O) t (glycosyl) x

where R2 has been selected from the group consisting of alkyl, alkylphenyl, hydroxyalkyl, hydroxyalkylphenyl and mixtures thereof, wherein the groups alkyl contain from about 10 to about 18, preferably from about 12 to about 14, atoms carbon; n is 2 or 3, preferably 2; t is from 0 to about 10, preferably 0; and x is about 1.3 to about 10, preferably from about 1.3 to about 3, most preferably about 1.3 to approximately 2.7. The glycosyl is preferably derived from the glucose. To prepare these compounds, alcohol is first formed or the alkyl polyethoxy alcohol and then it is done react with glucose or a source of glucose to form the glycoside (fixation in position 1). Glucosyl units Additional can be linked between position 1 and position 2, 3, 4 and / or 6 of the preceding glycosyl units, predominantly in the position 2.

Rust condensation products ethylene with a hydrophobic base formed by the condensation of oxide of propylene with propylene glycol are also suitable for use as additional non-ionic surfactant systems of the present invention The hydrophobic fraction of these compounds has preferably a molecular weight of about 1500 to approximately 1800 and presents insolubility in water. The adition of polyoxyethylene moieties at this hydrophobic portion tend to increase the water solubility of the molecule as a whole, and the character product liquid is preserved to the point where the Polyoxyethylene content is approximately 50% of total weight of the condensation product, which corresponds to a condensation of up to about 40 moles of ethylene oxide. Examples of compounds of this type include some of the surfactants Plurafac ™ LF404 and Pluronic ™, marketed by BASF

Also suitable for use as the non-ionic surfactant of the non-ionic surfactant system of the present invention are the condensation products of the oxide of ethylene and the product resulting from the reaction between oxide propylene and ethylenediamine. The hydrophobic rest of these products consists of the reaction product of ethylenediamine and oxide of excess propylene and generally has a molecular weight of approximately 2,500 to approximately 3,000. This hydrophobic residue is condensed with ethylene oxide to the extent that the product of condensation contains from about 40% to about 80% of polyoxyethylene by weight and has a molecular weight of approximately 5,000 to approximately 11,000. Examples of this Non-ionic surfactant type include some of the compounds Tetronic ™ commercials, marketed by BASF.

Preferred for use as the non-surfactant Ionic surfactant systems of the present invention are the condensates of poly (ethylene oxide) of alkylphenols, condensation products of primary aliphatic alcohols and secondary with about 1 to about 25 moles of ethylene oxide, alkylpolysaccharides, and mixtures thereof. Most preferred are C 8 -C 14 alkyl phenol ethoxylates having 3 to 15 ethoxy groups and alcohols C 8 -C 18 ethoxylated (preferably C 10 on average) having 2 to 10 ethoxy groups, and mixtures of the same.

Nonionic surfactants especially Preferred are acid polyhydroxyamide surfactants fatty formula

\delm{C}{\delm{\dpara}{O}}

---

\delm{N}{\delm{\para}{R ^{1} }}

--- Z,R2 ---

 \ delm {C} {\ delm {\ dpara} {O}} 

---

 \ delm {N} {\ delm {\ para} {R1}} 

--- Z,

wherein R 1 is H, or R 1 is C 1-4 hydrocarbyl, 2-hydroxyethyl, 2-hydroxy propyl or a mixture thereof, R2 is C 5-31 hydrocarbyl, and Z is a polyhydroxyhydrocarbyl with a chain linear hydrocarbyl with at least 3 attached hydroxyls directly thereto, or an alkoxylated derivative thereof. Preferably, R 1 is methyl, R 2 is a linear chain of C 11-15 alkyl or alkyl or alkenyl chain C 16-18 as, for example, cocoalkyl or mixtures thereof, and Z is a derivative of a sugar reducer such as glucose, fructose, maltose, lactose, in an amination reaction reductive

Anionic surfactants suitable for Use in the present invention are surfactants of the type linear alkyl benzene sulfonate and alkyl ester sulphonate included linear esters of carboxylic acids C 8 -C 20 (ie, fatty acids) sulphonated with gaseous SO3 according to "The Journal of the American Oil Chemists Society ", 52 (1975), pp. 323-329. Suitable starting materials would include fatty substances natural as those derived from tallow, palm oil, etc.

Alkyl ester sulfonate surfactants Preferred, especially for washing applications, comprise alkyl ester sulphonate surfactants with the formula structural:

\delm{C}{\delm{\para}{SO _{3} M}}

H ---

\uelm{C}{\uelm{\dpara}{O}}

--- OR^{4}R3 ---

 \ delm {C} {\ delm {\ para} {SO3 M}} 

H ---

 \ uelm {C} {\ uelm {\ dpara} {O}} 

--- OR 4

where R 3 is a hydrocarbyl C 8 -C 20, preferably an alkyl or a combination thereof, R 4 is a hydrocarbyl C 1 -C 6, preferably an alkyl or a combination thereof, and M is a cation that forms a salt Water soluble with alkyl ester sulfonate. The formation cations Suitable salts include metals such as sodium, potassium and lithium and substituted and unsubstituted ammonium cations, such as monoethanolamine, diethanolamine and triethanolamine. Preferably, R 3 is C 10 -C 16 alkyl and R 4 is methyl, ethyl or isopropyl. Especially preferred are the methyl ester sulphonates, wherein R 3 is alkyl C_ {10} -C_ {16}.

Other suitable anionic surfactants include alkyl sulfate surfactants that are salts or acids water soluble with the formula ROSO 3 M, where R is preferably a C 10 -C 24 hydrocarbyl, preferably an alkyl or hydroxyalkyl with a component C 10 -C 20 alkyl, more preferably a C 12 -C 18 alkyl or hydroxyalkyl, and M is H or a cation, for example an alkali metal cation (such as eg, sodium, potassium, lithium), or substituted ammonium or ammonium (by example cation of methylammonium, dimethylammonium and trimethylammonium and quaternary ammonium cation such as tetramethylammonium and dimethylpiperidinium cation and quaternary ammonium cation alkylamine derivatives such as ethylamine, diethylamine, triethylamine and mixtures thereof and the like). Typically, they are preferred C 12 {C} {16} alkyl chains for lower wash temperatures (e.g., lower than approximately 50 ° C) and alkyl chains C 16-18 for higher wash temperatures (e.g., higher than about 50 ° C).

In detergent compositions for washing clothes of the present invention may also include other anionic surfactants useful for detersive purposes. These can include salts (e.g., sodium, potassium, ammonium and salts of substituted ammonium such as monoethanolamine, diethanolamine salts and triethanolamine) of soap, alkanesulfonates C_ {8} -C_ {22} primary or secondary, C 8 -C 24 olefinsulfonates, acids sulfonated polycarboxylic acids prepared by sulfonation of the product pyrolysate of alkaline earth metal citrates, e.g. e.g., according to patent application GB-1,082,179, alkyl C 8 -C 24 polyglycol ether sulfates (which contain up to 10 moles of ethylene oxide); alkyl glycerol sulfonates, acyl glycerol fatty sulfonates, oleyl glycerol sulfates fatty, sulfuric ether of alkyl phenol ethylene oxide, sulphonates paraffin, alkyl phosphates, isethionates such as acyl isethionates, N-acyl taurates, alkyl succinamates and sulfosuccinates, sulphosuccinate monoesters (especially C 12 -C 18 saturated monoesters e unsaturated) and sulphosuccinate diesters (especially diesters C 6 -C 12 saturated and unsaturated), acyl sarcosinates, alkylpolysaccharide sulfates such as alkyl polyglucoside sulfates (non-ionic compounds not sulfated is described below), primary alkyl sulfates branched and alkyl polyethoxy carboxylates such as those of the formula RO (CH 2 CH 2 O) k -CH 2 COO-M + where R is a C 8 -C 22 alkyl, k is a integer from 1 to 10 and M is a soluble salt-forming cation. Too Resin acids and resin acids are suitable hydrogenates such as rosin, hydrogenated rosin and acids hydrogenated resins present in coniferous oil or derivatives thereof.

More examples are described in "Surface Active Agents and Detergents "(vol. I and II, by Schwartz, Perry and Berch). Some of said surfactants are also described in a manner General in US Patent 3,929,678, issued on 30 December 1975 to Laughlin et al., from column 23, line 58, to column 29, line 23.

If included, detergent compositions of washing according to the present invention typically comprise of about 1% to about 40%, preferably of about 3% to about 20%, by weight of surfactants Anionics of this type.

Especially preferred anionic surfactants include alkoxylated alkyl sulfate surfactants in the form of salts or water-soluble acids of formula RO (A) m SO3M, wherein R is an alkyl group or C 10 -C 24 hydroxyalkyl not substituted with a C 10 -C 24 alkyl component, preferably an alkyl or hydroxyalkyl C 12 -C 20, more preferably an alkyl or C 12 -C 18 hydroxyalkyl, is a unit ethoxy or propoxy, m is greater than zero, typically about 0.5 to about 6, more preferably of about 0.5 to about 3, and M is H or a cation that it can be for example a metal cation (for example sodium, potassium, lithium, calcium, magnesium, etc.), ammonium or a cation of substituted ammonium In the present invention both are contemplated ethoxylated alkyl sulfates such as propoxylated alkyl sulfates. Specific examples of substituted ammonium cations include methylammonium, dimethylammonium and trimethylammonium cations and cations of quaternary ammonium, such as cations of tetramethylammonium and dimethylpiperidinium as well as derivatives of alkylamines such as ethylamine, diethylamine, triethylamine, mixtures of The same and similar. Examples of surfactants are alkyl C 12 -C 18 polyethoxylate (1.0) sulfate (C 12 -C 18 E (1.0) M), the alkyl C 12 -C 18 polyethoxylate (2.25) sulfate (C 12 -C 18 E (2.25) M), the alkyl C 12 -C 18 polyethoxylate (3.0) sulfate (C 12 -C 18 E (3.0) M) and the alkyl C 12 -C 18 polyethoxylate (4.0) sulfate (C 12-C 18 E (4.0) M), where M Sodium and potassium are conveniently selected.

Cationic Detersive Surfactants suitable for use in washing detergent compositions according to the present invention are those that have a hydrocarbyl group of long chain. Examples of cationic surfactants of this type they include ammonium surfactants such as halides of alkyltrimethylammonium and surfactants with the formula:

[R 2 (OR 3) y] [R 4 (OR 3) y] 2 R 5 N + X-

wherein R 2 is an alkyl or alkyl benzyl group having from about 8 to about 18 carbon atoms in the alkyl chain, each R 3 is selected from the group consisting of -CH 2 CH 2 } -, -CH 2 CH (CH 3) -,
-CH 2 CH (CH 2 OH) -, -CH 2 CH 2 CH 2 -, and mixtures thereof; each R 4 is selected from the group consisting of C 1 -C 4 alkyl, C 1 -C 4 hydroxyalkyl, benzyl ring structures formed by the union of two R 4 groups , -CH 2 CHOH-CHOHCOR 6 CHOHCH 2 OH wherein R 6 is any hexose or hexose polymer with a molecular weight less than about 1000 and hydrogen when and is not 0; R 5 is the same as R 4 or is an alkyl chain wherein the total number of carbon atoms of R 2 plus R 5 is not more than about 18; each y is from 0 to about 10 and the sum of the values and is from 0 to about 15; and X is any compatible anion.

A suitable quaternary ammonium surfactant For the present invention it has the formula (I):

39

where R1 is a chain alkyl short (C6-C10) or alkylamidoalkyl of the formula (II):

40

and it's 2-4, preferably 3.

where R2 is H or an alkyl C1-C3,

where x is 0-4, preferably 0-2, most preferably 0,

where R3, R4 and R5 are the same or different and they can be a short chain (C1-C3) alkyl or alkoxylated alkyl of formula III,

where X - is a counterion, preferably a halide, for example chloride, or methylsulfate.

41

R6 is C 1 -C 4 and z is 1 or 2.

Quaternary ammonium surfactants Preferred are those defined by formula I where

R 1 is C 8, C 10 or mixtures of the themselves, x = o,

R 3, R 4 = CH 3 and R 5 = CH 2 CH 2 OH.

Highly cationic surfactants Preferred are water-soluble quaternary ammonium compounds useful in the present composition having the formula:

(i) R 1 R 2 R 3 R 4 N + X -

where R 1 is alkyl C 8 -C 16, each R 2, R 3 and R 4 they are, independently of each other, alkyl C 1 -C 4, hydroxyalkyl C 1 -C 4, benzyl and - (C 2 H 40) x H, where x has a value of 2 to 5, and X is a union. No more than one R 2, R 3 or R 4 must be benzyl

The preferred alkyl chain length for R 1 is C 12 -C 15, particularly when the alkyl group is a mixture of chain lengths derived from coconut or palm kernel oil or is derived by synthesis of accumulation of olefins or synthesis of OXO alcohols. The groups Preferred for R2, R3 and R4 are the methyl groups e hydroxyethyl and the anion X can be selected from halide ions, Metosulfate, acetate and phosphate.

Examples of quaternary ammonium compounds suitable with the formulas (i) of use in the present invention They are:

coconut trimethylammonium chloride or bromide;

Methyldihydroxyethylammonium chloride or bromide coconut;

decyltriethylammonium chloride;

chloride or bromide of decyldimethylhydroxyethylammonium;

C 12-15 chloride or bromide dimethyl hydroxyethyl ammonium;

dimethylhydroxyethylammonium chloride or bromide of coconut;

myristyltrimethylammonium methylsulfate;

chloride or bromide of lauryl dimethylbenzylammonium;

chloride or bromide of lauryl dimethyl (ethenoxy) 4-ammonium;

choline esters (compounds of formula (i) in where R_ {1} is

\delm{C}{\delm{\dpara}{O}}

--- C_{12-14} y R_{2}R_{3}R_{4} son metilo).alkyl CH2 --- CH_2 --- O ---

 \ delm {C} {\ delm {\ dpara} {O}} 

--- C 12-14 and R 2 R 3 R 4 are methyl).

dialkyl imidazolines [compounds of formula (i)].

Other cationic surfactants useful in the The present invention is also described in the patent US 4,228,044, issued October 14, 1980 to Change, and in the European patent application EP 000.224.

The cationic softening components of Typical fabrics include fabric softening active substances a water-insoluble quaternary ammonium base or its corresponding amine precursor, the most commonly used being chloride ammonium with double long alkyl chain or methyl sulfate.

Of these, the preferred cationic softeners They include the following:

one)

disebo-dimethylammonium chloride (DTDMAC);

2)

dihydrogenated tallow dimethylammonium chloride;

3)

tallow dimethylammonium dihydroxy methylsulfate;

4)

distearyl dimethylammonium chloride;

5)

dioleyl dimethylammonium chloride;

6)

dipalmityl hydroxyethyl methyl ammonium chloride;

7)

stearyl benzyl dimethylammonium chloride;

8)

tallow trimethylammonium chloride;

9)

hydrogenated tallow trimethylammonium chloride;

10)

alkyl chloride C 12 -14 hydroxyethyl dimethylammonium;

eleven)

C 12-18 alkyl chloride dihydroxyethyl methyl ammonium;

12)

di (stearoyloxyethyl) chloride dimethylammonium (DSOEDMAC);

13)

chloride di (tallow-oxy-ethyl) dimethylammonium;

14)

distilled methylsulfate imidazolinium;

fifteen)

methylsulfate 1- (2-seboylamidoethyl) -2-seboyl imidazolinium

Ammonium compounds have been present biodegradable quaternaries as alternatives to chlorides Long alkyl double chain ammoniums and methyl sulfates traditionally used. These quaternary ammonium compounds contain long chain alkylo groups interrupted by functional groups such as carboxyl groups. The fabric softener products and compositions that they contain are described in numerous publications such as, e.g. eg EP-A-0.040.562 and EP-A-0.239.910.

Quaternary ammonium compounds and amine precursors of the present invention have the formulas (I) or (II) following:

42

where Q is selected from -O-C (O) -, -C (O) -O-, -O-C (O) -O-, -NR 4 -C (O) -, -C (O) -NR 4 -;

R1 is (CH 2) n -Q-T 2 or T3;

R2 is (CH 2) m -Q-T 4 or T 5 or R 3;

R 3 is alkyl C 1 -C 4 or hydroxyalkyl C 1 -C 4 or H;

R 4 is H or alkyl C 1 -C 4 or hydroxyalkyl C 1 -C 4;

T 1, T 2, T 3, T 4, T 5 are, independently of each other, alkyl or alkenyl C 11 -C 22;

n and m are integers from 1 to 4; Y

X - is an anion compatible with the softener. Among the non-limiting examples of compatible anion Chloride or methylsulfate are included with the softener.

The alkyl or alkenyl chain T1, T 2, T 3, T 4, T 5 must contain at least 11 atoms of carbon and preferably at least 16 carbon atoms. The chain can be linear or branched. Sebum is an adequate source and economical of long chain alkyl and alkenyl compounds. The compounds wherein T 1, T 2, T 3, T 4, T 5 represent the mixture of tallow long chain compounds are the especially preferred.

Specific examples of ammonium compounds Quaternary suitable for use in aqueous compositions Fabric softeners of the present invention include:

one)

chloride N, N-di (seboyl-oxy-ethyl) -N, N-dimethyl ammonium;

2)

methylsulfate N, N-di (seboyl-oxy-ethyl) -N-methyl, N- (2-hydroxyethyl) ammonium;

3)

chloride N, N-di (2-seboyl-oxy-2-oxo-ethyl) -N, N-dimethyl ammonium;

4)

chloride N, N-di (2-seboyl-oxy-ethylcarbonyl-oxy-ethyl) -N, N-dimethyl ammonium;

5)

chloride N- (2-seboyl-oxy-2-ethyl) -N- (2-seboyl-oxy-2-oxo-ethyl) -N, N-dimethyl;

6)

chloride N, N, N-tri (seboyl-oxy-ethyl) -N-methyl ammonium;

7)

chloride N- (2-seboyl-oxy-2-oxo-ethyl) -N- (seboyl-N, N-dimethyl-ammonium Y

8)

chloride 1,2-diseboyl-oxy-3-trimethylammonium propane;

and mixtures of any of the products previous.

If included in it, the compositions laundry detergents according to the present invention contain typically from 0.2% to about 25%, preferably from about 1% to about 8%, by weight of surfactants Cationics of this type.

Laundry detergent compositions of the present invention may also contain surfactants ampholytic, hybrid ion and semipolar as well as non-surfactants ionic and / or anionic different from those already described herein memory.

Ampholytic surfactants are also suitable for use in washing detergent compositions of clothing of the present invention. These surfactants can Generically described as aliphatic derivatives of amines secondary or tertiary, or aliphatic derivatives of amines secondary or tertiary heterocyclics in which the radical Aliphatic can be a linear or branched chain. One of the aliphatic substituents contains at least about 8 atoms carbon, typically from about 8 to about 18 carbon atoms, and at least one contains an anionic group Water solubilizer, for example carboxy, sulphonate, sulfate. See US Patent 3,929,678, issued to Laughlin et al. on December 30, 1975, in column 19, lines 18-35, for examples of surfactants ampholytic

If included in the present invention, the washing detergent compositions according to it consist typically from 0.2% to about 15%, preferably from about 1% to about 10%, by weight of surfactants amphibitics of this type.

Hybrid ion surfactants are also suitable for use in detergent compositions for washing clothes. These surfactants can be broadly described as derivatives of secondary and tertiary amines, derivatives of amines secondary and tertiary heterocyclic or derivatives of compounds of quaternary ammonium, quaternary phosphonium or tertiary sulfonium. See US Patent 3,929,678, issued to Laughlin et al. on December 30, 1975, in column 19, line 38 until the column 22, line 48, for examples of ionic surfactants hybrid.

If included, detergent compositions of wash according to the present invention typically contain from 0.2% to about 15%, preferably about 1% at approximately 10%, by weight of hybrid ion surfactants of this kind.

Semi-polar non-ionic surfactants are a special category of non-ionic surfactants that include oxides of water soluble amines containing an alkyl moiety of about 10 to about 18 carbon atoms, and 2 residues selected from the group consisting of alkyl groups and hydroxyalkyl groups containing from about 1 to about 3 carbon atoms; phosphine oxides soluble in water containing an alkyl moiety of about 10 to approximately 18 carbon atoms and 2 moieties selected from group consisting of alkyl groups and hydroxyalkyl groups that they contain from about 1 to about 3 atoms of carbon; and water soluble sulfoxides containing a residue alkyl of about 10 to about 18 atoms of carbon and a remainder selected from the group consisting of residues alkyl and hydroxyalkyl from about 1 to about 3 carbon atoms

Nonionic Detergent Surfactants semipolar include amine oxide type surfactants of formula:

\uelm{N}{\uelm{ \uparrow }{0}}

(R^{5})2R 3 (OR 4) x

 \ uelm {N} {\ uelm {\ uparrow} {0}} 

(R 5) 2

where R 3 is a group alkyl, hydroxyalkyl, or alkylphenyl or mixtures thereof that it contains from about 8 to about 22 atoms of carbon; R 4 is an alkylene or hydroxyalkylene group that contains from about 2 to about 3 carbon atoms or mixtures thereof; x is 0 to about 3; and every R 5 is an alkyl or hydroxyalkyl group containing about 1 to about 3 carbon atoms or a group poly (ethylene oxide) containing about 1 to approximately 3 groups of ethylene oxide. R 5 groups they can be linked together, for example by an atom of oxygen or nitrogen, to form a structure of ring.

These amine oxide type surfactants include, in particular, alkyl oxides C 10 -C 18 -dimethyl-amine and alkoxy oxides C 8 -C 12 -ethyl-hydroxy-ethyl-amine.

When included in the composition, the cleaning compositions of the present invention typically comprise from 0.2% to about 15%, preferably from about 1% to about 10%, by weight of these semi-polar nonionic surfactants.

The laundry detergent composition of the present invention may also comprise a surfactant auxiliary selected from the group of primary amines or tertiary.

The primary amines suitable for use in the Present invention include amines according to the formula R1 NH2 where R_ {1} is an alkyl chain C 6 -C 12, preferably C_ {6} -C_ {10} or R 4 X (CH 2) n, X is -O-, -C (O) NH- or -NH-, R4 is an alkyl chain C 6 -C 12, n is from 1 to 5 and preferably 3. The alkyl chains R1 can be linear or branched and they can be interrupted by up to 12, preferably less than 5, ethylene oxide residues.

Preferred amines according to the formula plus Above herein are n-alkylamines. Amines suitable for use according to the present invention may be selected from 1-hexylamine, 1-octylamine, 1-decylamine and laurylamine Other preferred primary amines include C8-C10 Oxypropylamine, octyl-oxypropylamine, 2-ethylhexyl oxypropylamine, lauryl-amidopropylamine and amidopropylamine.

Tertiary amines suitable for use in Present invention include tertiary amines having the formula R 1 R 2 R 3 N where R1 and R2 are alkyl chains C_ {1} -C_ {8} or

\uelm{C}{\uelm{\para}{R _{5} }}

H --- O)_{x}H--- (CH_ {2} ---

 \ uelm {C} {\ uelm {\ para} {R5}} 

H --- O) x H

R_ {3} is or an alkyl chain C 6 -C 12, preferably a chain C 6 -C 10 alkyl or R 3 is R 4 X (CH 2) n, where X is -O-, -C (O) NH- or -NH-, R4 is an alkyl chain C_ {4} -C_ {12}, n is between 1 and 5, preferably 2-3. R 5 is H or an alkyl group C_ {1} -C_ {2} and x is between 1 and 6.

R 3 and R 4 can be alkyl chains linear or branched; R 3 alkyl chains may be interrupted by up to 12, preferably less than 5, residues of ethylene oxide.

Preferred tertiary amines are R 1 R 2 R 3 N where R 1 is an alkyl chain C6-C12, R2 and R3 are C1-C3 alkyl or

\uelm{C}{\uelm{\para}{R _{5} }}

H --- O)_{x}H--- (CH_ {2} ---

 \ uelm {C} {\ uelm {\ para} {R5}} 

H --- O) x H

where R5 is H or CH3 and x = 1-2.

Amidoamines are also preferred with the formula:

\uelm{C}{\uelm{\dpara}{O}}

--- NH --- (CH_{2})_{n} --- N --- (R_{2})_{2}R_ {1} ---

 \ uelm {C} {\ uelm {\ dpara} {O}} 

--- NH --- (CH 2) n --- N --- (R 2) 2

where R 1 is an alkyl C 6 -C 12; n is 2-4,

n is preferably 3; R 2 and R 3 is C_ {1} -C_ {4}

The most preferred amines herein invention include 1-octylamine, 1-hexylamine, 1-decylamine, 1-dodecylamine, oxy C8-10 propylamine, N coconut 1-3-diaminopropane, coconut alkyldimethylamine, lauryl dimethylamine, lauryl bis (hydroxyethyl) mine, bis (hydroxyethyl) coconut amine, lauryl amine 2 mol propoxylated, 2 mol propoxylated octylamine, lauryl amidopropyl dimethylamine, amidopropyl C8-10 dimethylamine and amidopropyl C10 dimethylamine

The most preferred amines for use in compositions according to the present invention are 1-hexylamine, 1-octylamine, 1-decylamine, 1-dodecylamine. They are especially desirable n-dodecyldimethylamine and coconut bis and hydroxy ethylalkylamine ethoxylated 7 times, lauryl-amidopropylamine and coconut amidopropylamine

Bleaching agent

Laundry detergent compositions of the present invention may also comprise an agent bleach such as hydrogen peroxide, PB1, PB4 and percarbonate with a particle size of 400-800 micrometers. These bleaching agent components may contain one or more oxygen bleaching agents and, according to the agent chosen bleach, one or more bleach activators. When oxygen bleaching compounds are present typically they will be present at levels of approximately 1% at approximately 25%

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The bleaching component for use herein invention can be any of the useful bleaching agents for detergent compositions, including bleach oxygen releasers as well as others known in the art. He bleaching agent suitable for the present invention may be a bleaching agent activated or not activated.

A class of bleaching agent releasing oxygen for use in the present invention encompasses the agents bleaches based on percarboxylic acid and salts of same. Suitable examples of this class of agents include the magnesium monoperoxyphthalate hexahydrate magnesium salt meta-chloro perbenzoic acid, acid 4-nonylamino-4-oxoperoxybutyric and diperoxidedecanedioic acid. Bleaching agents of this type are described in US Pat. No. 4,483,781, the US-740,446, EP-0.133.354 and the US 4,412,934. Bleaching agents Especially preferred also include the acid 6-nonylamino-6-oxoperoxicaproproic  as described in US Patent 4,634,551.

Another category of bleaching agents that they can be used encompassing halogen bleaching agents. Examples of hypohalite bleaching agents include for example trichloroisocoanuric acid, sodium dichloroisocyanurates and potassium and N-chloro-alkanesulfonamides and N-bromo-alkanesulfonamides. These materials are normally added in quantities of 0.5-10% by weight of the finished product, preferably 1-5% by weight.

Hydrogen peroxide releasing agents they can be used together with bleaching effect activators such such as tetraacetylethylenediamine (TAED), the nonanoyloxybenzene sulphonate (NOBS, described in US 4,412,934), 3, 5, -trimethylhexanoloxybenzenesulfonate (ISONOBS, described in EP 120,591), pentaacetylglucose (PAG) or the phenolsulfonate ester of acid N-nonanoyl-6-aminocaproic acid (NACA-OBS, described in WO94 / 28106), which are perhydrolysed to form a peracid that acts as the active bleach providing a better bleaching effect. Acylated citrate esters are also suitable activators. such as those described in the codependent patent application EP 624 154 and the acyclic imide bleach activators asymmetric of the following formula, as described in the Procter & co-pending patent application WO 98/04664 Gamble:

43

where R_ {1} is an alkyl group C 7 -C 13 straight or branched chain, saturated or unsaturated, R2 is an alkyl group C 1 -C 8, straight or branched chain, saturated or unsaturated and R 3 is an alkyl group C 1 -C 4 straight or branched chain, saturated or not saturated.

Useful bleaching agents, including peroxyacids and bleaching systems comprising activators of bleach and peroxy bleaching compounds for use in the detergent compositions according to the invention are described in our codependent requests WO 95/10592, WO 97/00937, WO95 / 27772, WO95 / 27773, WO95 / 27774 and WO95 / 27775.

Hydrogen peroxide can also be present if an enzymatic system is added (i.e. an enzyme and a substrate for it) capable of generating hydrogen peroxide at the beginning or during the washing and / or rinsing process. These Enzymatic systems are described in patent application EP 537 381, filed October 9, 1991.

Catalysts that contain metal for your use in bleaching compositions include catalysts that they contain cobalt such as pentaamine acetate salts and cobalt (III) and catalysts containing manganese such as described in EPA 549 271; EPA 549 272; EPA 458 397; US 5,246,621; EPA 458 398; US 5,194,416 and US 5,114,611. A bleaching composition that comprises a peroxy compound, a bleach catalyst that contains manganese and a chelating agent is described in the application Patent 94870206.3.

Bleaching agents other than agents Oxygenated bleaches are also known in the art and They can be used in the present invention. A type of agent Non-oxygen bleach of special interest are the photoactivated bleaching agents such as phthalocyanines from sulfonated zinc and / or aluminum. These materials can be deposited in the substrates during the washing process. Irradiation with light in the presence of oxygen, such as when hanging clothes on outside in daylight, causes the activation of phthalocyanine of sulphonated zinc and, consequently, the bleaching of the substrate. A preferred zinc phthalocyanine and a photoactivated bleaching process It is described in US Patent 4,033,718. Typically, the detergent compositions will contain about 0.025% at about 1.25%, by weight, of zinc phthalocyanine sulphonated

Strengthening detergency system

Preferably, the detergent compositions of laundry of the present invention may also comprise a detergent builder additive, more preferably a zeolite, a sodium tripolyphosphate and / or a laminar silicate. It has been surprisingly discovered that these compositions provide better cleaning capacity, especially in stains cosmetics and food, and advantages of better release of the dirt.

Any detergency builder system Conventional is suitable for use in the present invention, including aluminosilicate products, silicates, polycarboxylates, alkyl succinic acid or alkenyl succinic acid and fatty acids, products such as ethylenediamine tetraacetate, diethylenetriamine-pentamethylene acetate, sequestrants of metal ions such as aminopolyphosphonates, especially acid ethylenediamine tetramethylene phosphonic acid diethylenetriamine-pentamethylene phosphonic. In the present invention reinforcing agents can also be used phosphate type detergency.

Additives strengthening detergency suitable may be an inorganic ion exchange product, generally an inorganic product of hydrated aluminosilicate, more especially a synthetic hydrated zeolite like zeolite hydrated A, X, B, HS or MAP.

Other detergency builder material suitable inorganic is laminar silicate, for example SKS-6 ™ (Hoechst). SKS-6 is a crystalline laminar silicate consisting of sodium silicate (Na 2 Si 2 O 5).

Suitable polycarboxylates containing a carboxy group include lactic acid, glycolic acid and derivatives ether thereof as described in the Belgian patents 831,368, 821,369 and 821,370. Polycarboxylates containing two carboxy groups include the water soluble salts of succinic acid, Malonic acid, (ethylenedioxy) diacetic acid, maleic acid, acid diglycolic, tartaric acid, tartronic acid and fumaric acid, as well as the carboxylate ether described in German patents 2,446,686, and 2,446,687 and US 3,935,257 and the sulfinil carboxylates described in Belgian patent 840,623. The polycarboxylates containing three carboxy groups include, in particular, citrates, aconitrates and water soluble citraconate as well as succinate derivatives such as carboxymethyloxysuccinates described in GB-1,379,241, lactoxysuccinates described in Dutch patent 7205873, and the materials of oxypolycarboxylate such as 2-oxa-1,1,3-propane tricarboxylates described in GB-1,387,447.

Polycarboxylates containing four groups carboxy include the oxidisuccinates described in BG-1,261,829, the 1,1,2,2-ethane tetracarboxylates, 1,1,3,3-propane-tetracarboxylates and the 1,1,2,3-propane-tetracarboxylates. Polycarboxylates containing sulfo substituents include the sulphosuccinate derivatives described in the patents GB-1,398,421 and GB-1,398,422 and in the US Patent 3,936,448, and pyrolysed citrates sulfonates described in GB-1,082,179, in so much so that polycarboxylates containing phosphonium substituents It is described in patent GB-1,439,000.

Alicyclic polycarboxylates and heterocyclic include cyclopentane-cis, cis, cis-tetracarboxylates, cyclopentadiene-pentacarboxylates, 2,3,4,5-tetrahydro-furan - cis, cis, cis-tetracarboxylates, 2,5-tetrahydro-furan -cis - dicarboxylates, 2,2,5,5-tetrahydrofuran - tetracarboxylates, 1,2,3,4,5,6-hexane -hexacarboxylates and carboxymethyl derivatives of alcohols polyhydric agents such as sorbitol, mannitol and xylitol. The aromatic polycarboxylates include melitic acid, acid pyromellitic and phthalic acid derivatives described in the GB-1,425,343 patent.

Of the aforementioned, the Preferred polycarboxylates are hydroxycarboxylates that they contain up to three carboxy groups per molecule and more in particular Citrates

The additive systems reinforcing the Preferred detergency for use in the present compositions include a mixture of a detergent builder additive of water insoluble aluminosilicate such as zeolite A or a silicate laminar (SKS-6) and a carboxylate chelating agent Water soluble like citric acid. Other additive systems Preferred detergency builders include a mixture of a insoluble aluminosilicate detergency builder in water like zeolite A and a carboxylate chelating agent Water soluble like citric acid. Additive systems Preferred detergency builder for use in detergent liquid compositions of the present invention are those soaps and polycarboxylates.

Other detergency reinforcing materials which may be part of the detergency booster system of Use in granulated compositions include inorganic materials such as alkali metal carbonates, bicarbonates, silicates and organic materials such as organic phosphonates, phosphonates of aminopolyalkylene and aminopolycarboxylates.

Other suitable water-soluble organic salts are homo polymeric or copolymeric acids or their salts, in the that the polycarboxylic acid comprises at least two radicals carboxyl separated from each other by no more than two carbon atoms. In patent GB-A-1,596,756 is describe polymers of this type. Examples of these salts are the polyacrylates with molecular weight of 2,000-5,000 and their copolymers with maleic anhydride, these copolymers having a molecular weight of 20,000 to 70,000, especially about 40,000

The detergent builder salts are normally included in amounts of 5% to 80% by weight of the composition, preferably from 10% to 70% and usually from 30% to 60% by weight.

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Conventional Detergent Enzymes

Laundry detergent compositions they may comprise, in addition to the enzyme mannanase, one or more enzymes that provide advantages of cleaning capacity, care of tissues and / or disinfection.

Such enzymes include enzymes selected from cellulases, hemicellulases, peroxidases, proteases, gluco-amylases, amylases, xylanases, lipases, phospholipases, esterases, cutinases, pectinases, keratanases, reductases, oxidases, phenoloxidases, lipoxygenases, ligninases, pululanasas, tanasas, pentosanasas, malanasas, β-glucanases, arabinosidases, hyaluronidases, Chondroitinases, lacases or mixtures thereof.

A preferred combination is a composition. laundry detergent that has a combination of enzymes Conventional applicable as protease, amylase, lipase, cutinase and / or cellulase with one or more wall degradation enzymes plant cell.

Suitable proteases are the subtilisin that are obtained from specific strains of B. subtilis and B. licheniformis (subtilisin BPN and BPN '). A suitable protease is obtained from a Bacillus strain that has a maximum activity in the pH range of 8-12, and is developed and marketed as ESPERASE® by Novo Industries A / S of Denmark, hereinafter "Novo". The preparation of this enzyme and analogous enzymes is described in patent GB-1,243,784 to Novo. Other suitable proteases include ALCALASE®, DURAZYM® and SAVINASE® de Novo and MAXATASE®, MAXACAL®, PROPERASE® and MAXAPEM® (genetically engineered protein from Maxacal) from Gist-Brocades. Proteolytic enzymes also encompass modified bacterial serine proteases, such as those described in application EP 251 446, filed on April 28, 1987 (especially pages 17, 24 and 98), and which is referred to herein as "Protease B "and in application EP-199,404, Venegas, filed on October 29, 1986, which refers to a modified bacterial serine proteolytic enzyme which is referred to as" Protease A "herein. The protease referred to herein as "protease C", which is a variant of an alkaline Bacillus serine protease in which lysine replaces arginine at position 27, tyrosine replaces valine at position 104, is suitable. serine replaces asparagine at position 123 and alanine replaces threonine at position 274. Protease C is described in EP 451 244 corresponding to WO 91/06637, published May 16, 1991. Genetically modified variants, especially protease C, they are also included herein.

A preferred protease, hereinafter "protease D" is a variant of carbonyl hydrolase that has an amino acid sequence that is not found in nature and that is derived from a carbonyl hydrolase precursor by substitution of a different amino acid with a plurality of residues amino acid at a position in said carbonyl hydrolase equivalent to the +76 position, preferably also together with one or more positions of amino acid residues equivalent to those selected from the group consisting of +99, +101, +103, +104, +107, +123, +27, +105, +109, +126, +128, +135, +156, +166, +195, +197, +204, +206, +210, +216, +217, +218 , +222, +260, +265 and / or +274 according to the Bacillus amiloliquefaciens subtilisin numbering, according to WO95 / 10591 and in the patent application of C. Ghosh et al., "Bleaching Compositions Comprising Protease Enzymes" WO 95 / 10592, filed on October 13, 1994. A carbonyl hydrolase variant of the protease described in WO95 is also suitable. / 10591 having an amino acid sequence derived by substitution of a plurality of amino acid residues substituted in the precursor enzyme at the +210 position together with one or more of the following residues: +33, +62, +67, +76, + 100, +101, +103, +104, +107, +128, +129, +130, +132, +135, +156, +158, +164, +166, +167, +170, +209, +215, +217, +218 and +222, where the numbered position corresponds to the natural subtilisin of Bacillus amiloliquefaciens or equivalent amino acid residues in other carbonyl hydrolases or subtilisins such as subtilisin Bacillus lentus (codependent patent application WO 98/55634 , filed June 4, 1997).

They are also suitable for the present invention. the proteases described in EP-251 applications 446 and WO-91/06637, the BLAP® protease described in WO91 / 02792 and its variants described in WO 95/23221.

See also a high pH protease from Bacillus sp . NCIMB 40338 described in WO 93/18140 A granted to Novo. WO 92/03529 A, granted to Novo, describes enzymatic detergents comprising protease, one or more additional enzymes and a reversible protease inhibitor. If desired, there is a protease with reduced adsorption and increased hydrolysis as described in WO 95/07791 issued to Procter & Gamble. In WO 94/25583 granted to Novo a trypsin-like recombinant protease for suitable detergents according to the present invention is described. Other suitable proteases are described in EP 516 200 issued to Unilever.

Proteolytic enzymes are incorporated into the laundry detergent compositions of the present invention at a level of 0.0001% to 2%, preferably 0.001% to 0.2%, more preferably from 0.005% to 0.1%, of pure enzyme by weight of the composition.

Cellulases useful in the present invention include bacterial and fungal cellulases. Preferably, they will have an optimum pH of 5 to 12 and a specific activity greater than 50 CEVU / mg (cellulose viscosity unit). Suitable cellulases are described in US Pat. No. 4,435,307, Barbesgoard et al., J61078384 and WO96 / 02653, which describe a fungal cellulase produced, respectively, from Humicola insolens, Trichoderma, Thielavia and Sporotrichum . EP 739 982 describes cellulases isolated from a new species of Bacillus. Suitable cellulases are also described in GB-A-2,075,028; GB-A-2,095,275; DE-OS-2,247,832 and WO95 / 26398.

Examples of these cellulases are cellulases obtained from a strain of Humicola insolens ( Humicola grisea var. Thermoidea ), in particular the Humicola strain DSM 1800.

Other suitable cellulases are Humicola insolens cellulases having a molecular weight of approximately 50 KDa, an isoelectric point of 5.5 and containing 415 amino acids and a \43 kD endoglucanase derived from Humicola insolens , DSM 1800, which has cellulase activity ; a preferred endoglucanase component has the amino acid sequence described in PCT application WO 91/17243. Also suitable are cellulases EGIII cellulases of Trichoderma longibrachiatum described in WO94 / 21801, Genencor, published on September 29, 1994. Especially suitable cellulases are cellulases that have advantages for color care. Examples of these cellulases are the cellulases described in European patent application EP 495 257, published on November 6, 1991 (Novo). Carezyme and Celluzyme (Novo Nordisk A / S) are especially useful. See also WO91 / 17244 and WO91 / 21801. Other suitable cellulases with tissue care and / or cleaning properties are described in WO96 / 34092, WO96 / 17994 and WO95 / 24471.

Said cellulases are normally incorporated in the laundry detergent composition at levels of 0.0001% at 2% pure enzyme by weight of the washing detergent composition of clothes.

Peroxidase enzymes are used together with oxygen sources, e.g. eg, percarbonate, perborate, persulfate, hydrogen peroxide, etc. and with a phenolic substrate as a molecule bleach improver. They are used to "whiten the solution ", that is, to prevent the transfer of dyes or pigments removed from the substrates during the operations of wash to other substrates in the wash solution. Enzymes peroxidase are known in the art and include, for example, horseradish peroxidase, ligninase and haloperoxidase such as chloro-peroxidase and the bromine peroxidase. Detergent compositions that contain peroxidase are described, for example, in the application PCT International WO 89/099813, WO89 / 09813 and in the application for European patent EP 540 784, published on November 6, 1991. The enzyme lacasa is also suitable.

The improvers are generally understood at a level of 0.1% l 5% by weight of the total composition. The Preferred enhancers are phentiazine and phenoxyasin substituted, 10-phenothiazinepropionic acid (PPT), the acid 10-ethylphenothiazin-4-carboxylic (EPC), 10-phenoxazinpropionic acid (POP) and 10-methylphenoxazine (described in WO 94/12621), the syringates (C3-C5 alkyl substituted syringates) and the substituted phenols. Sodium percarbonate or perborate are preferred sources of hydrogen peroxide.

Said peroxidases are normally incorporated. in the laundry detergent composition at levels of 0.0001% at 2% pure enzyme by weight of the washing detergent composition of clothes.

Other preferred enzymes that may be included in the laundry detergent compositions of the present invention include lipases. Lipase enzymes suitable for use in detergent include those produced by microorganisms of the Pseudomonas group, such as, for example, Pseudomonas stutzeri ATCC 19,154, as described in GB-1,372,034. Suitable lipases include those that exhibit a positive immunological cross-reaction with the lipase antibody, produced by the Pseudomonas fluorescent IAM 1057 microorganism. This lipase is marketed by Amano Pharmaceutical Co. Ltd., Nagoya, Japan, under the trade name lipase P "Amano", hereinafter referred to as "Amano-P". Other suitable commercial lipases include Amano-CES, lipases ex Chromobacter viscosum , for example Chromobacter viscosum var . NRRLB 3673 lipolyticum from Toyo Jozo Co., Tagata, Japan; Chromobacter viscosum lipases from US-Biochemical Corp., USA and Disoynth Co., Holland, and lipases of Pseudomonas gladioli . Especially suitable lipases are lipases such as M1 Lipase® and Lipomax® (Gist-Brocades) and Lipolase® and Lipolase Ultra® (Novo) which have been found to be very effective when used in combination with the compositions of the present invention. Also suitable are the lipolytic enzymes described in EP 258 068, WO 92/05249 and WO 95/22615 by Novo Nordisk and in WO 94/03578, WO 95/35381 and WO 96/00292 by Unilever.

Cutinases are also suitable [EC 3.1.1.50] which can be considered as a special type of lipase, to Know lipases that do not require an interface activation. The Addition of cutinases to detergent compositions has been described by example in WO-A-88/09367 (Genencor); WO 90/09446 (Plant Genetic System) and WO 94/14963 and WO 94/14964 (Unilever).

Lipases and / or cutinases are incorporated normally in the laundry detergent composition at levels from 0.0001% to 2% pure enzyme by weight of the detergent composition of laundry.

Amylases (α and / or β) may be included to remove carbohydrate stains. In WO94 / 02597, granted to Novo Nordisk A / S on February 3, 1994, are described detergent compositions incorporating mutant amylases. Watch also WO95 / 10603, granted to Novo Nordisk A / S on April 20, 1995. Other amylases known for use in compositions detergents include α-amylases and β-amylases. The α-amylases are known in the art and they include those described in US Patents 5,003,257; EP 252,666; WO / 91/00353; FR 2,676,456; EP 285,123; EP 525,610; EP 368,341; and GB-1,296,839 (Novo). Other amylases suitable are amylases with improved stability described in WO94 / 18314, published on August 18, 1994, and WO96 / 05295, granted on February 22, 1996 to Genencor, and variants of amylase that has an additional modification in the precursor immediately marketed by Novo Nordisk A / S, described in WO 95/10603, published in April 1995. Also suitable are Amylases described in EP 277 216, WO95 / 26397 and WO96 / 23873 (all of Novo Nordisk).

Product Examples Purafect Ox Am® commercial α-amylases from Genencor and Termamyl®, Ban®, Fungamyl® and Duramyl®, all of them marketed by Novo Nordisk A / S Denmark. WO95 / 26397 describes other suitable amylases: α-amylases characterized by having a specific activity of at least 25% exceeding the specific activity of Termamyl® at a range of temperature of 25ºC to 55ºC and with a pH value in the range of 8 to 10, measured by the activity test of Phadebas® α-amylase. They are adequate variants of the above enzymes, described in WO96 / 23873 (Novo Nordisk). Other amylolytic enzymes with better properties of activity level and with a combination of thermostability and a higher level of activity are described in WO95 / 35382.

Amylolytic enzymes are incorporated into the laundry detergent compositions of the present invention at a level of 0.0001% to 2%, preferably from 0.00018% to 0.06%, more preferably from 0.00024% to 0.048%, of pure enzyme by weight of the composition.

The enzymes mentioned above may be of any suitable origin, such as vegetable, animal, Bacterial, fungal or yeast. The origin can also be mesophilic or extremophilic (psychrophilic, psychotropic, thermophilic, barophilic, alkalophilic, acidophilic, halophilic, etc.). Can be used purified or unpurified forms of these enzymes. Currently It is a common practice to modify wild-type enzymes by protein / genetic engineering techniques to optimize your performance in laundry detergent compositions of the invention. For example, variants can be designed to increase the compatibility of the enzyme with respect to the ingredients of Common use in these compositions. Alternatively, the variant It can be designed in such a way that the optimum pH, stability of the bleach or chelator, catalytic activity and the like of the Enzyme is suitable for the particular cleaning application.

In particular, special attention should be given to oxidation sensitive amino acids for the stability of bleach and in the case of surface charges for surfactant compatibility. The point can be modified. isoelectric of these enzymes replacing some amino acids loaded so that, p. eg, an increase in the isoelectric point may help improve compatibility with surfactants anionic Enzyme stability can also be improved creating, p. eg, additional salt bridges and causing the Calcium binding sites increase chelator stability. Special attention should be given to cellulases since most of the cellulases have separate binding domains (CBD). They can alter the properties of these enzymes by modifying these domains

Such enzymes are normally incorporated into the laundry detergent composition at levels from 0.0001% to 2% of pure enzyme by weight of the washing detergent composition of clothes. Enzymes can be added as independent ingredients separately (pellets, granules, stabilized liquids, etc. that they contain an enzyme) or as mixtures of two or more enzymes (e.g. cogranulates).

Other suitable detergent ingredients that can be added are the oxidation eliminators of enzymes that are described in the codependent patent application EP-92870018.6, filed on January 31, 1992. Examples of these enzyme oxidation scavengers are the tetraethylene ethoxylated polyamines.

In WO 9307263 A and WO 9307260 A, of Genencor International, WO 8908694 A, de Novo, and the patent US-3,553,139, granted to McCarty et al. on 5 of January 1971, different materials are also described enzymatic and media for incorporation into the compositions synthetic detergents Enzymes are also described in the patent. US 4,101,457, granted to Place et al. on 18 of July 1978 and US Patent 4,507,219, granted to Hughes on March 26, 1985. Enzymatic materials useful for liquid detergent formulations and for the incorporation of themselves in such formulations are described in the patent US 4,261,868, issued to Hora et al. April 14th of 1981. Enzymes for use in detergent can be stabilized through different techniques. In the patent US-3,600,319, granted to Gedge et al. on 17 of August 1971 and EP-199,405 and EP-200,586, granted to Venegas on October 29 1986, techniques for the stabilization of enzymes Enzyme stabilization systems are described also, for example, in US Patent 3,519,570. A species of Bacillus, AC13, useful that produces proteases, xylanases and cellulases are described in WO 9401532 A de Novo.

Advantages for the care of colors and fabrics

You can also include technologies that They provide a type of color protection advantage. Examples of these technologies are metal catalysts for conservation of colors. These metal catalysts are described in the EP-92870181.2 codependent patent application. Dye fixatives, polyolefin dispersion for prevent wrinkles and improve water absorbency, perfumes and polymers with amine functionality (PCT / US97 / 16546) for the treatment of color care and perfume permanence are other examples of technologies for the care of colors and of the tissues.

In detergent compositions for washing clothes according to the present invention can also be incorporated fabric softening agents. These agents can be of type inorganic or organic. Examples of inorganic softeners are the smectite clays described in GB-A-1,400,898 and in the USP 5,019,292. Organic fabric softeners include amines water insoluble tertiary as described in GB-A1.514.276 and EP-B0.011.340 and its combination with mono quaternary ammonium salts C12-C14 described in EP-B-0.026.527 and EP-B-0.026.528 and chain diamides long as described in EP-B-0.242.919. Other ingredients Useful fabric softener systems include materials high molecular weight poly (ethylene oxide) as described in EP-A-0.299.575 and 0.313.146.

Smectite clay levels are normally from 2% to 20%, more preferably from 5% to 15% in weight, adding the product as a dry mixed component to the rest of the formulation Organic fabric softeners such as materials based on non-water-soluble tertiary amines or amides Long double chain are incorporated at levels of 0.5% to 5% in weight, usually 1% to 3% by weight, while the materials of high molecular weight poly (ethylene oxide) and materials Water-soluble cationics are added in amounts of 0.1% to 2%, usually from 0.15% to 1.5% by weight. These materials are added normally to the spray dried portion of the composition, although in some cases it may be more appropriate to add them as a mixture of dry mixed particles or spray them in liquid form cast on the solid components of the composition.

Chelating agents

Laundry detergent compositions of the present invention may also optionally contain one or more iron and / or manganese chelating agents. Chelating agents of this type can be selected from the group consisting of aminocarboxylates, aminophosphonates, aromatic chelating agents polyfunctionally substituted and their mixtures, all of them as They are defined below. Without pretending to impose any theory, it think that the advantage of these materials is due, in part, to their exceptional ability to separate iron and manganese ions from washing solutions by forming soluble chelates.

The aminocarboxylates useful as agents Optional chelators include ethylenediaminetetracetates, N-hydroxyethylethylene diamino triacetates, nitrilotriacetates, ethylenediamine tetraproprionates, triethylene tetraamine hexacetates, diethylene triamino pentaacetates, and ethanoldiglycins, alkali metal salts, ammonium, and ammonium substituted thereof and mixtures thereof

Amino phosphonates are also suitable for their use as chelating agents in the compositions of the invention when at least low levels of total phosphorus are allowed in laundry detergent compositions and include ethylenediamine tetrakis (methylene phosphonates) such as DEQUEST ™. Preferably these amino phosphonates do not contain alkyl or alkenyl groups with more than approximately 6 carbon atoms.

Aromatic chelating agents Polyfunctionally substituted are also useful in Compositions of this invention. See the patent US 3,812,044, issued May 21, 1974 to Connor et al. Preferred compounds of this type in acid form they are dihydroxydisulfobenzenes, such as 1,2-dihydroxy-3,5-disulfobenzene.

A preferred biodegradable chelator for use in the present invention is ethylenediamine disuccinate ("EDDS"), especially the [S, S] isomer as described in the patent US 4,704,233, issued November 3, 1987 to Hartman and Perkins.

The compositions of the present invention they can also contain water-soluble salts (or acid form) of the methyl glycine di-acetic acid (MGDA) as a chelator or detergency builder co-additive useful with, for example, insoluble detergency builder additives such as zeolites, laminar silicates and the like.

If used, these chelating agents generally will comprise from about 0.1% to about 15% by weight of the laundry detergent compositions of the present invention More preferably, if used, the agents chelators will comprise from about 0.1% to about 3.0% by weight of such compositions.

Soap suppressant

Another optional ingredient is an antifoam, illustrated by silicones and mixtures of silica-silicone Usually silicones are represented by alkylated polysiloxane materials while silica is normally used in finely divided forms such as, p. eg, aerogels and xerogels of silica and hydrophobic silicas of different types. These materials can be incorporated in the form of particles in which the suds suppressor is advantageously incorporated releasably into a vehicle waterproof detergent, water soluble or water dispersible, basically no surfactant. Alternatively, the suppressor of Soaps can dissolve or disperse in a vehicle liquid and applied by spraying in one or more of additional components

A regulator of silicone soaps Preferred is described in the patent, granted to Bartollota et al., US-3 933 672. Other suds suppressors particularly useful are the suds suppressors of self-emulsifying silicone, described in the patent application German DTOS 2 646 126, published on April 28, 1977. An example Compound of this type is DC-544, marketed by Dow Corning, which is a copolymer of siloxane glycol. A regulator of soaps Especially preferred is the suds suppressor system comprising a mixture of silicone oils and 2-alkyl alkanoles. The Suitable 2-alkyl alkanols are the 2-butyl octanol sold under trademark Isofol 12 R.

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This suds suppressor system is described in EP 593 841 codependent patent application, filed on November 10, 1992.

Silicone soaps regulators Especially preferred are described in the patent application EP 573 699 codependent. Such compositions may comprise a silicone / silica mixture together with non-porous smoke silica such like Aerosil®.

The defoamers described above are normally used at levels of 0.001% to 2% by weight, preferably from 0.01% to 1% by weight, of the composition.

Others

Other components may be used in the laundry detergent compositions such as hangers of dirt, agents to release dirt, brighteners optical, abrasive, bactericidal, fogging inhibitors, dyes and / or encapsulated or non-encapsulated perfumes.

Especially suitable encapsulating products they are water-soluble capsules consisting of a polysaccharide plate and polyhydroxy compounds such as those described in GB 1,464,616. Other suitable encapsulating water-soluble products comprise dextrin derived from esters of non-gelatinized acid starch substituted dicarboxylic such as those described in US 3,455,838. These ester-derived dextrins they are prepared, preferably, from these starches as corn waxy, sorghum, sago, tapioca and potato. Suitable examples of said encapsulating materials include N-Lok, manufactured by National Starch. The encapsulating material of N-Lok consists of a modified corn starch and glucose. The starch has been modified by adding groups monofunctional substitutes such as acid anhydride octenyl succinic.

Anti-redeposition agents and suspensions of Suitable dirt of the present invention include derivatives of cellulose such as methyl cellulose, carboxymethyl cellulose e hydroxyethylcellulose and homo polymeric polycarboxylic acids or copolymers or their salts. Polymers of this type include the polyacrylates and copolymers of maleic anhydride-acrylic acid listed above as detergency builder additives, as well as copolymers of maleic anhydride with ethylene, ether of methylvinyl or methacrylic acid, constituting maleic anhydride at least 20 moles percent of the copolymer. These materials are normally used in amounts of 0.5% to 10% by weight, plus preferably from 0.75% to 8%, most preferably from 1% to 6%, by weight of the composition.

Preferred optical brighteners are of anionic type as, for example, 4,4'-bis- (2-diethanolamino-4-anilino-s-triazin-6-ylamino) stilbene-2,2'-disulfonate disodium 4,4'-bis- (2-morpholino-4-anilino-s-triazin-6-ylamino-stilbene-2,2'-disulfonate disodium 4,4'-bis- (2,4-dianilino-s-triazin-6-ylamino) stilbene-2,2'-disulfonate disodium 4 ', 4' '- bis- (2,4-dianilino-s-triazin-6-ylamino) stilbene-2-sulphonate monosodium, 4,4'-bis- (2-anilino-4- (N-methyl-N-2-hydroxyethylamino) -s-triazin-6-ylamino) stilbene-2,2'-disulfonate disodium 4,4'-bis- (4-phenyl-2,1,3-triazol-2-yl) -tylbeno-2,2'-disulfonate disodium 4,4'bis (2-anilino-4- (1-methyl-2-hydroxyethylamino) -s-triazin-6-ylamino) stilbene-2,2'-disulfonate disodium 2 (stilbil-4 '' - (naphtho-1 ', 2', 4.5) -1,2,3-triazol-2 '' - sulfonate sodium and 4,4'-bis (2-sulfoestyryl) biphenyl. Highly preferred brighteners are brighteners. specific ones described in EP 753 567.

Other useful polymer products are the polyethylene glycols, especially those of molecular weight of 1,000 to 10,000, more preferable those of 2,000 to 8,000 and with maximum preference of approximately 4,000. These are used to levels from 0.20% to 5%, and more preferably from 0.25% to 2.5%, in weight. These polymers and homopolymeric polycarboxylate salts and copolymers mentioned above are effective in improving the conservation of whiteness, the deposition of ashes from the tissue and the ability to clean clay stains, of nature protein and oxidizable in the presence of metal impurities of transition.

Conventional dirt release polymers

Preferably, the detergent compositions of laundry of the present invention will comprise another polymer for the release of conventional dirt. This composition provides better cleaning and releasing ability of dirt The polymer for dirt release suitable is polyester with anion protected ends and conventional form copolymers or terpolymers of terephthalic acid with ethylene glycol and / or propylene glycol units in different arrangements such as polypropylene diethoxylated terephthalate. Examples of these polymers are described in the patents of common assignment US-4116885 and US-4711730 and EP-0 272 033. A particular preferred polymer according to EP-A-0 272 033 has the formula

(CH 3 (PEG) 43) 0.75 (POH) 0.25 [T-PO) 2.8 (T-PEG) 0.4 ] T (PO-H) 0.25 ((PEG) 43 CH 3) 0.75

where PEG is - (OC_ {2} H4} O-, PO is (OC_ {3} H_ {0}) and T is (pcOC_ {6} H4 {CO}).

Polyesters are also very useful modified as random copolymers of dimethyl terephthalate, dimethyl sulfoisophthalate, ethylene glycol and 1-2 propanediol, the terminal groups that consist first of sulfobenzoate and secondly of ethylene glycol and / or monoesters propanediol The goal is to get a polymer terminally protected at both ends by sulfobenzoate groups; "primarily", in the present context, most of these copolymers according to the present invention will have protected ends by sulfobenzoate groups. However, some copolymers do not have fully protected terminals, so that your groups terminals may consist of ethylene glycol monoester and / or propane-1-2-diol, "secondarily" comprising these variants.

The polyesters selected according to the present invention contain about 46% by weight of acid dimethylterephthalic, approximately 16% by weight of propane-1,2 diol, approximately 10% by weight of ethylene glycol, approximately 13% by weight of acid dimethylsulfobenzoic acid and approximately 15% by weight of acid sulfoisophthalic, and have a molecular weight of approximately 3,000 Polyesters and their method of preparation are described in detail in EPA 311 342.

It is well known in the art that chlorine free in tap water quickly deactivates enzymes included in the detergent compositions. Therefore, the use of chlorine scavengers such as perborate, ammonium sulfate, sulphite sodium or polyethyleneimine at a level greater than 0.1%, by weight of the total composition in the formulas will provide a better Stability during washing of detergent enzymes. Compositions comprising a chlorine remover are found described in European patent application EP 553 607 published on January 31, 1992.

Alkoxylated polycarboxylates, such as prepared from polyacrylates, are useful in this invention to provide a fat removal capacity additional. This type of materials is described in WO-91/08281 and in the document PCT-90/01815 on p. 4 and next Chemically, these Materials comprise polyacrylates with an ethoxy side chain for every 7-8 acrylate units. The chains laterals are of the formula - (CH 2 CH 2 O) m (CH 2) n CH 3, where m is 2-3 and n is 6-12. The side chains are attached with ester to the "chain main "polyacrylate providing a type structure "combined" polymer. Molecular weight may vary, but typically it is in the range of about 2,000 to Approximately 50,000 Alkoxylated polycarboxylates of this type they can comprise from about 0.05% to about 10% in weight, of the compositions of this invention.

Dispersants

Laundry detergent compositions of the present invention may also contain dispersants. You go out Suitable water-soluble organic are homopolymeric acids or copolymers or their salts in which the polycarboxylic acid it comprises at least two carboxyl radicals separated from each other by no More than two carbon atoms. In the patent GB-A-1,596,756 are described polymers of this type. Examples of these salts are the polyacrylates with molecular weight of 2,000-5,000 and their copolymers with maleic anhydride, these copolymers having a molecular weight from 1,000 to 100,000.

Especially, copolymers of acrylate and methylacrylate such as 480N that have a weight molecular weight of 4000, at a level of 0.5-20% by weight of the composition in laundry detergent compositions of The present invention.

The compositions of the invention may contain a lime soap peptizer compound, which has preferably a dispersing capacity of lime soap (LSDP) as defined below, not more than 8, preferably not more than 7 and most preferably no more than 6. The compound Lime soap peptizer is preferably present at a level from 0% to 20% by weight.

A quantification of the effectiveness of a limestone soap peptizer is obtained by the ability limestone soap dispersant (LSDP) that is determined by limestone soap dispersant test according to an article by H.C. Borghetty and C.A. Bergman, J. Am. Oil. Chem. Soc., Vol. 27, p. 88-90, (1950). This dispersion test method of calcareous soap is widely used by technicians in this field of art and reference, for example, in the following articles; W.N. Linfield, Surfactant science Series, vol. 7, p. 3; W.N. Linfield, Tenside surf. det., vol. 27, p. 159-163, (1990); and M.K. Nagarajan, W.F. Masler, Cosmetics and Toiletries, vol. 104, p. 71-73, (1989). LSDP is the weight ratio (in%) between the agent dispersant and the sodium oleate required to disperse the limestone soap deposits formed by 0.025 g of sodium oleate in 30 ml of 333 ppm water of CaCo3 (Ca: Mg = 3: 2) equivalent of hardness.

Surfactants with a good action Lime soap peptizer will include certain amine oxides, betaines, sulfobetaines, alkyl ethoxy sulfates and alcohols ethoxylated.

Among the illustrative surfactants that have an LSDP of no more than 8 for use in the present invention is include dimethyl amine oxide C 16 -C 18, the alkyl C 12 -C 18 ethoxy sulfates with a medium grade of ethoxylation of 1-5, especially the surfactant C 12 -C 15 alkyl ethoxy sulfate with a grade ethoxylation of 3 (LSDP = 4) and ethoxylated alcohols C 14 -C 15 with an average degree of ethoxylation 12 (LSDP = 6) or 30, marketed under the registered trademarks Lutensol A012 and Lutensol A030, respectively, by BASF GmbH.

Polymeric limestone soap peptizers suitable for use in the present invention are described in the article by M.K. Nagarajan, W.F. Masler, which is in Cosmetics and Toiletries, vol. 104, p. 71-73, (1989).

They can also be used as peptizers of chalky soap hydrophobic bleaches such as 4- [N-octanoyl-6-aminohexanoyl] benzene sulfonate, 4- [N-nonanoyl-6-aminohexanoyl] benzene sulfonate, 4- [N-decanoyl-6-aminohexanoyl] benzene sulfonate and mixtures thereof; and nonanoyloxy benzene sulfonate together with bleaching formulations hydrophilic / hydrophobic.

Inhibition of dye transfer

Laundry detergent compositions of the present invention may also include compounds for inhibit the transfer of one tissue to another of the dyes dissolved and suspended present during washing operations of colored fabrics.

Polymeric agents transfer inhibitors dyes

Laundry detergent compositions according to the present invention they also comprise from 0.001% to 10%, preferably from 0.01% to 2%, more preferably from 0.05% to 1%, in weight of dye transfer inhibitors polymeric Said agents that inhibit the transfer of polymeric dyes are normally incorporated into the compositions laundry detergents to inhibit the transfer of dyes from colored fabrics to fabrics washed with the same. These polymers have the property of adsorbing or of complex with fugitive dyes from colored tissues before they can be fixed on others washed products

Polymeric transfer inhibitors Especially suitable dyes are N-oxide polymers of polyamine, copolymers of N-vinyl pyrrolidone and N-vinylimidazole, polyvinylpyrrolidone polymers, polyvinyloxazolidones and polyvinylimidazoles or mixtures of the same.

The addition of these polymers also improves the Enzyme yield according to the invention.

a) Polyamine N-oxide polymers

Suitable polyamine N-oxide polymers for use in the invention contain units with the following structural formula:

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44

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where P is a polymerizable unit, the group R-N-O may be attached to or form part of the polymerizable unit, or a combination of both.

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Four. Five

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R are aliphatic, aliphatic ethoxylated groups, heterocyclic or alicyclic aromatics or any combination of the same to which the group's nitrogen can be fixed N-O or where the group's nitrogen N-O is part of these groups.

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The N-O group can be represented by the following general structures:

46

where R1, R2 and R3 are groups aliphatic, aromatic, heterocyclic or alicyclic or combinations of these, x o / y y o / y z is 0 or 1 and where the nitrogen of the N-O group may be attached to or be part of these groups

The N-O group can be part of the polymerizable unit (P) or it can be attached to the chain polymeric principal, or a combination of both.

Suitable polyamine N-oxides in which the N-O group is part of the polymerizable unit they comprise the polyamine N-oxides in which R is selected from aliphatic, aromatic, alicyclic or heterocyclic groups.

A class of said polyamine N-oxides it comprises the group of polyamine N-oxides in which the nitrogen from the N-O group is part of the R group. Preferred polyamine N-oxides are those in which R is a heterocyclic group such as, for example, pyridine, pyrrole, imidazole, pyrrolidine, piperidine, quinoline, acridine and derivatives of same.

Another class of said polyamine N-oxides comprises the group of polyamine N-oxides in which nitrogen from group N-O is linked to group R.

Other suitable polyamine N-oxides are those polyamine oxides in which the N-O group is attached to the polymerizable unit.

The preferred class of these N-oxides of Polyamine are the polyamine N-oxides with the general formula (I) wherein R is an aromatic, heterocyclic or alicyclic group in the that the nitrogen of the N-O functional group is part of said group R.

Examples of these classes are the oxides of polyamine in which R is a heterocyclic compound as, by example, pyridine, pyrrole, imidazole and derivatives thereof.

Another preferred class of these N-oxides of polyamine are the polyamine oxides with the general formula (I) in where R are aromatic, heterocyclic or alicyclic groups in the that the nitrogen of the N-O functional group is attached to these groups R.

Examples of these classes are the oxides of polyamine where R groups can be aromatic as phenyl.

Any main chain can be used polymer as long as the amine oxide polymer formed is water soluble and have transfer inhibitory properties of dyes They are examples of polymeric main chains Suitable: polyvinyl, polyalkylene, polyesters, polyethers, polyamides, polyimides, polyacrylates and mixtures thereof.

The amine N-oxide polymers of the present invention typically have an amine ratio: amine N-oxide from 10: 1 to 1: 1,000,000. However, the amount of amine oxide groups present in the amine polyoxide polymer by a suitable copolymerization or by an adequate degree of N-oxidation. Preferably, the relationship amine: Amine N-oxide is from 2: 3 to 1: 1,000,000, more preferably from 1: 4 to 1: 1,000,000 and most preferably from 1: 7 to 1: 1,000,000. The polymers of the present invention actually encompass random or block copolymers in which one type of monomer is an amine N-oxide and the other type of monomer may or may not be a Amine N-Oxide The amine oxide unit of the N-oxides of polyamine has a PKa <10, preferably a PKa <7 and more preferably a PKa <6.

Polyamine oxides can be obtained with almost any degree of polymerization. The degree of polymerization It is not critical if the material has the desired properties in in terms of water solubility and suspension power of dyes

Typically, the average molecular weight is in the range of 500 to 1,000,000; preferably from 1,000 to 50,000, more preferably 2,000 to 30,000 and with maximum preference of 3,000 to 20,000.

b) Copolymers of N-vinyl pyrrolidone and N-vinylimidazole

N-vinylimidazole polymers and N-vinyl pyrrolidone used herein invention have an average molecular weight of 5,000-1,000,000 and preferably 5,000 to 200,000

The most preferred polymers for use in laundry detergent compositions according to the present invention comprise a polymer selected from copolymers of N-vinylimidazole N-vinyl pyrrolidone wherein said polymer has an average molecular weight in the range from 5,000 to 50,000, more preferably from 8,000 to 30,000 and most preferably 10,000 to 20,000.

The average molecular weight range is determined by light diffusion as described in Barth H.G. Y Mays J.W. Chemical Analysis vol. 113, "Modern Methods of Polymer Characterization ".

The copolymers of N-vinylimidazole N-vinyl pyrrolidone especially preferred have a molecular weight range average of 5,000 to 50,000; more preferably from 8,000 to 30,000; most preferably 10,000 to 20,000.

The copolymers of N-vinylimidazole and N-vinyl pyrrolidone characterized by having said average molecular weight range provide excellent inhibition ability dye transfer and do not adversely affect the cleaning performance of formulated detergent compositions with them.

The copolymers of N-vinylimidazole N-vinyl pyrrolidone according to the present invention they have a molar ratio of N-vinylimidazole with respect to N-vinyl pyrrolidone from 1 to 0.2, more preferably from 0.8 to 0.3, most preferably from 0.6 to 0.4.

c) Polyvinylpyrrolidone

Laundry detergent compositions of the present invention can also use polyvinylpyrrolidone ("PVP") with an average molecular weight of about 2,500 to about 400,000, preferably of about 5,000 to about 200,000, more preferably from about 5,000 to about 50,000, and with maximum preference of about 5,000 to about 15,000. The Suitable polyvinylpyrrolidones are marketed by ISP Corporation, New York, NY and Montreal, Canada, with the names of PVP product K-15 (molecular weight in viscosity of 10,000), PVP K-30 (average molecular weight of 40,000), PVP K-60 (average molecular weight of 160,000), and PVP K-90 (average molecular weight of 360,000). Other suitable polyvinylpyrrolidones are those marketed by BASF Cooperation that include Sokalan ™ HP 165 and Sokalan ™ HP 12; and polyvinylpyrrolidones known to the expert in the field of detergents (see, for example, EP-A-262,897 and EP-A-256,696).

d) Polyvinyloxazolidone

Laundry detergent compositions of the present invention can also use polyvinyloxazolidone as a transfer inhibiting agent for polymeric dyes. Said polyvinyloxazolidones have a weight average molecular from about 2,500 to about 400,000, preferably from about 5,000 to about 200,000, more preferably from about 5,000 to about 50,000, and most preferably about 5,000 to Approximately 15,000

e) Polyvinylimidazole

Laundry detergent compositions of the present invention they can also use polyvinylimidazole as a polymeric dye transfer inhibitor. These polyvinylimidazoles have an average of approximately 2,500 to about 400,000, preferably about 5,000 to about 200,000, more preferably from about 5,000 to approximately 50,000, and most preferably approximately 5,000 to approximately 15,000.

f) Crosslinked polymers

Crosslinked polymers are polymers whose Main chains are interconnected to a certain degree. These links can be chemical or physical in nature, possibly with active groups in the main chain or in the ramifications. Crosslinked polymers have been described in Journal of Polymer Science, volume 22, p. 1035-1039.

In one embodiment, the crosslinked polymers are prepare so that they form a three-dimensional rigid structure that can trap dyes in the pores formed by the three-dimensional structure In another embodiment, the polymers reticulates trap dyes due to swelling. These crosslinked polymers are described in the patent application. codependent EP 719 856.

Washing method

The compositions of the invention can be used in virtually any washing or cleaning method, including soaking methods, pretreatment methods and methods with rinsing stages in which a separate rinse enhancing composition.

The process described in the present invention comprises contacting tissues with a wash solution of the usual form and exemplified below. The process of the Invention is properly carried out during the cleaning process. The cleaning method is preferably executed between 5 ° C and 95 ° C, especially between 10ºC and 60ºC. The pH of the treatment solution It is preferably 7 to 12.

The following examples are included for illustrate the compositions of the present invention although without necessarily limit or otherwise define the scope of the invention. In laundry detergent compositions the Enzyme levels are expressed in pure enzyme by weight of the total composition and, unless otherwise indicated, detergent ingredients are expressed by weight of the composition total. The abbreviations of the components herein They have the following meaning:

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THE

: Sodium alkyl sulfonate C 11-13 linear benzene.

TAS

: Sebum sodium alkyl sulfate.

CxyAS

: Sodium Alkylsulfate C_ {1x} -C_ {1y}.

CxySAS

: (2,3) alkyl C 1x-C 1y sodium sulfate secondary.

CxyEz

: Condensed Primary Alcohol C_ {1x} -C_ {1y} predominantly linear with a z moles average of {} \ hskip0.1cm oxide ethylene.

CxyEzS

: Condensed sodium alkylsulfate C_ {1x} -C_ {1y} with an average of z moles of ethylene oxide.

QAS

: R 2 .N + (CH 3) 2 (C 2 H 4 OH) with R 2 = C 12 {C 14}.

QAS 1

: R 2 .N + (CH 3) 2 (C 2 H 4 OH) with R2 = C_ {8} -C_ {11}.

APA

: Propyl dimethyl amine amido C_ {8-10}.

Soap

: Linear alkyl sodium carboxylate derived from an 80/20 mixture of tallow and coconut fatty acids.

Nonionic

: C 13 -C 15 fatty alcohol mixed ethoxylated / propoxylated with an average degree of ethoxylation of 3.8 and {} \ hskip0.1cm an average degree of propoxylation of 4,5.

Neodol 45-13

: Alcohol C14-C15 linear ethoxylated primary, marketed by Shell Chemical CO.

STS

: Sodium toluenesulfonate.

CFAA

: Alkyl C 12 -C 14 N-methyl glucamide.

TFAA

: Rent C_ {16} -C_ {18} N-methyl glucamide.

TPKFA

: C 12 -C 14 fatty acids derived from the primary distillation of crude oil.

Silicate

: Amorphous sodium silicate (ratio SiO2: Na2O = 1.6-3.2).

Metasilicate

: Sodium Metasilicate (SiO2 ratio: Na2O   = 1.0).

Zeolite A

: Hydrated sodium aluminosilicate of formula Na 12 (AlO 2 SiO 2) 12. 27H_2 or that it has a size {} \ hskip0.1cm of primary particles in the range of 0.1 to 10 micrometers (weight expressed as anhydrous substance}.

Na-SKS-6

: Crystalline laminar silicate of formula δ-Na_ {2} Si_ {2} O_ {5}

Citrate

: Trisodium citrate dihydrate with one activity of 86.4% and a size distribution of partí - {} \ hskip0,1cm cula from 425 to 850 micrometers

Citric

: Anhydrous citric acid.

Borate

: Sodium Borate

Carbonate

: Anhydrous sodium carbonate with a size of particles between 200 and 900 micrometers.

Baking soda

: Anhydrous sodium acid carbonate with a particle size distribution between 400 and 1200 {} \ micrometre.

Sulfate

: Anhydrous sodium sulfate.

{} \ Hskip0.4cm sulfate magnesium

{} \ hskip0.5cm: Sulfate anhydrous magnesium

STPP

: Sodium tripolyphosphate.

TSPP

: Tetrasodium pyrophosphate.

MA / AA

: Random copolymer with a 4: 1 ratio of acrylate: maleate and an average molecular weight from {} \ hskip0.1cm approximately 70,000 to 80,000.

MA / AA 1

: Random copolymer with a 6: 4 ratio of acrylate: maleate and an average molecular weight of {} \ hskip0,1cm approximately 10,000

AA

: Sodium polyacrylate polymer with a weight Average molecular weight of 4,500.

PA30

: Polyacrylic acid with a molecular weight average of approximately 4,500 to 8,000.

480N

: Random copolymer with a 7: 3 ratio of acrylate: methacrylate and a molecular weight average {} \ hskip0.1cm of approximately 3,500.

Poligel / carbopol

: High weight crosslinked polyacrylates molecular.

PB1

: Anhydrous sodium perborate monohydrate of formula Nominal NaBO_2 .H2 {2}.

PB4

: Sodium perborate tetrahydrate of formula Nominal NaBO2 .3H2 {2 O.H2 {O2}.

Percarbonate

: Anhydrous sodium percarbonate of nominal formula 2Na2CO3 .3H2 {O2}.

NaDCC

: Sodium dichloroisocyanurate.

TAED

: Tetraacetylethylenediamine.

NOBS

: Nonanoyloxybenzene sulphonate in salt form Sodium

NACA-OBS

: (6-nonamidocaproyl) sulfonate  Oxybenzene

DTPA

: Diethylenetriamine pentaacetic acid.

HEDP

: 1,1-hydroxyethane acid diphosphonic

DETPMP

: Diethyltriamino penta (methylene) phosphonate, marketed by Monsanto with the name commercial {} \ hskip0,1cm Dequest 2060.

EDDS

: Acid ethylenediamino-N, N'-disuccinic, isomer (S, S) in the form of its sodium salt

MnTACN

: 1,4,7-trimethyl-1,4,7-triazacyclononane   Manganese

Bleach photoactivated

: Zinc Phthalocyanide sulfonated encapsulated in dextrin soluble polymer.

Photoactivated bleach one

: Sulfonated aluminum phthalocyanide encapsulated in dextrin soluble polymer.

PAAC

: Pentaamino acetate salt cobalt (III).

Paraffin

: Paraffin oil marketed under the name Winog 70 by Wintershall.

Nabz

: Sodium benzoate.

BzP

: Benzoyl peroxide.

Mananasa

: Bacillus agaradherens Mannanase, MCIMB 40482.

Protease

: Proteolytic enzyme distributed with the name commercial of Savinase, Alcalase, Durazym by {} \ hskip0,1cm Novo Nordisk A / S, Maxacal, Maxapem marketed by Gist-Brocades and proteases described {} \ 0,15 in WO91 / 06637 and / or WO95 / 10591 and / or EP 251 446.

Amylase

: Amylolytic enzyme marketed under the name Purafact Ox Am® described in WO 94/18314, {} \ hskip0.1cm WO96 / 05295 marketed by Genencor; Termamyl®, Fungamyl® and Duramyl®, all of them {} \ hskip0,1cm marketed by Novo Nordisk A / S and those described in WO95 / 26397.

Lipase

: Lipolytic enzyme marketed under the name commercial Lipolase, Lipolase Ultra by Novo {} \ hskip0,1cm Nordisk A / S and Lipomax by Gist-Brocades.

Cellulase

: Cellulite enzyme marketed with trade names Carezyme, Celluzyme and / or Endo - {} \ hskip0,1cm lase by Novo Nordisk A / S.

CMC

: Sodium carboxymethylcellulose.

PVP

: Polyvinyl polymer, with a molecular weight 60,000 average.

PVNO

: Polyvinylpyridine N-oxide, with a weight Average molecular weight of 50,000.

PVPVI

: Copolymer of vinylimidazole and vinylpyrrolidone, with an average molecular weight of 20,000.

Brightener 1

: 4,4'-bis (2-sulfoestiryl) biphenyl   disodium

Brightener 2

: 4,4'-bis (4-anilino-6-morpholino-1,3,5-triazin-2-yl)   stilbene-2,2'-disulfonate disodium

Defoamer type silicone

: Foam controller polydimethylsiloxane with copolymer of siloxane-oxyalkylene as {} \ hskip0.1cm dispersing agent with a relationship between said controller of foam and said dispersing agent {} \ 10cm from 10: 1 a 100: 1

 Suppressor of suds

{} \ hskip0.1cm: 12% of silicone / silica, 18% stearyl alcohol, 70% starch in granulated form.

Opacifying

: Aqueous mixture of monostyrene latex, marketed by BASF AG under the brand registered {} \ Lytron 621.

SRP 1

: Polyesters with ends protected with anion.

SRP 2

: Poly (1,2 propylene terephthalate) polymer short block diethoxylate

QEA

: bis ((C 2 H 5 O) (C 2 H 4 O) n) (CH 3) -N + - C 6 H 12 -N + - (CH 3) bis ((C 2 H 5 O) - (C 2 H 4 O)) n, where n = from 20 to 30

PEI

: Polyethyleneimine such as PEI 1800 E7, PEI 1200 E7, PEI 1200 E7 quaternized, PEI 600 E20 as {} \ 0.1cm is described in WO97 / 42288.

SCS

: Sodium Cumensulfonate.

HMWPEO

: high weight poly (ethylene oxide) molecular.

PEG

: Polyethylene glycol, with a molecular weight of x.

PEO

: poly (ethylene oxide), with a molecular weight 5,000 average.

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Example 1

The according to the present invention were prepared following high density detergent compositions for washed:

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Example 2

The according to the present invention were prepared following granulated detergent compositions for washing Particular utility in machine wash conditions in Europe:

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Example 3

The according to the present invention were prepared following detergent compositions of particular utility in machine wash conditions in Europe:

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Example 4

The according to the present invention were prepared following granulated detergent compositions:

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108

Example 5

The according to the present invention were prepared following detergent compositions that do not contain bleach Especially useful for washing colored clothes:

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Example 6

The according to the present invention were prepared following detergent compositions:

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Example 7

The according to the present invention were prepared following granulated detergent compositions:

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Example 8

The according to the present invention were prepared following detergent compositions:

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Example 9

The according to the present invention were prepared following detergent compositions:

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Example 10

The following formulations were prepared liquid detergents according to the present invention (the levels are express in parts by weight and enzymes in pure enzyme):

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Example 11

The following formulations were prepared liquid detergents according to the present invention (the levels are express in parts by weight and enzymes in pure enzyme):

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Example 12

The according to the present invention were prepared following liquid detergent compositions (levels are express in parts by weight and enzymes in pure enzyme):

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Example 13

The according to the present invention were prepared following liquid detergent compositions (levels are express in parts by weight and enzymes in pure enzyme):

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Example 14

The according to the present invention were prepared following granulated detergent compositions for fabrics with a "softening during washing" action:

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Example 15

The according to the present invention were prepared following detergent compositions in the form of a soap bar (levels are expressed in parts by weight and enzymes in enzyme pure):

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Example 16

The according to the present invention were prepared following detergent compositions with additives:

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(1. GENERAL INFORMATION:

APPLICANT: NAME: The Procter \ amp {1} Gamble Company STREET: One Procter \ amp {1} Gamble Square CITY: Cincinnati, OHIO COUNTRY: USA POSTAL CODE: 45202

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TITLE OF THE INVENTION: Detergent compositions comprising a mannanase and a polymer for the release of the dirt.

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NUMBER OF SEQUENCES: 6

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LEGIBLE FORM FOR THE COMPUTER: TYPE OF SUPPORT: Floppy COMPUTER: PC compatible with IBM OPERATING SYSTEM: PC-DOS / MS-DOS SOFTWARE: PatentIn Release No. 1.0 Version 1.25 (EPO)

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SEC. one

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CHARACTERISTICS OF THE SEQUENCE: LENGTH: 1407 pairs of bases TYPE: acid nucleic FILAMENT: simple TOPOLOGY: linear

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TYPE OF MOLECULE: DNA genomic ORIGINAL SOURCE

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CHARACTERISTIC: NAME / KEY: CDS POSITIONS: 1-1482

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DESCRIPTION OF THE SEQUENCE: SEC. one

47

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SEC. 2

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CHARACTERISTICS OF THE SEQUENCE: LENGTH: 493 amino acids KIND: amino acid TOPOLOGY: linear

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TYPE OF MOLECULE: protein

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DESCRIPTION OF THE SEQUENCE: SEC. 2

48

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SEC.3

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CHARACTERISTICS OF THE SEQUENCE: LENGTH: 1407 pairs of bases TYPE: acid nucleic FILAMENT: simple TOPOLOGY: linear

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TYPE OF MOLECULE: DNA genomic

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DESCRIPTION OF THE SEQUENCE: SEC. 3

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SEC. 4

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CHARACTERISTICS OF THE SEQUENCE: LENGTH: 468 amino acids KIND: amino acid TOPOLOGY: linear

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TYPE OF MOLECULE: protein

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DESCRIPTION OF THE SEQUENCE: SEC. 4

fifty

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SEC. 5

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CHARACTERISTICS OF THE SEQUENCE: LENGTH: 1029 pairs of bases TYPE: acid nucleic FILAMENT: simple TOPOLOGY: linear

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TYPE OF MOLECULE: DNA genomic

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DESCRIPTION OF THE SEC SEQUENCE 5

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52

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SEC. 6

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CHARACTERISTICS OF THE SEQUENCE: LENGTH: 363 amino acids KIND: amino acid TOPOLOGY: linear

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TYPE OF MOLECULE: protein

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DESCRIPTION OF THE SEQUENCE: SEC. 6

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Claims (11)

1. A laundry detergent composition comprising a mannanase enzyme, a polymer for the release of dirt for cotton fabrics of polyethyleneimine type and at less an additional detergent component. 2. A laundry detergent composition according to claim 1, wherein said mannanase is present at a level of 0.0001% to 2%, preferably 0.0005% to 0.5%, more preferably from 0.001% to 0.1%, of pure enzyme by weight of the total composition 3. A laundry detergent composition according to claims 1-2, wherein the Dirt release polymer for cotton fabrics polyethyleneimine type is comprised at a level from 0.0001% to 20%, preferably from 0.001% to 15%, more preferably from 0.01% to 10%. 4. A laundry detergent composition according to claims 1-3, wherein the Dirt release polymer for cotton fabrics Polyethyleneimine type has the following formula: 55 which has a polyamine formula modified V (n + 1)} W_ {m} Y_ {n} Z or a main chain polyamine according to formula: 56 which has a polyamine formula modified V (n-k + 1)} W_m Y_ {n} Y 'k Z, in where k is less than or equal to n, said main chain having polyamine before modification a molecular weight greater than approximately 200 daltons, in where

i)

 the V units are terminal units that have the formula:

57

ii)

the W units are main chain units that have the formula:

58

iii)

the Y units are branching units that have the formula:

59

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iv)

the Z units are terminal units that have the formula:

60

where the R units binding to the main chain are selected from the group consisting of C2-C12 alkylene, C4-C12 alkenylene, hydroxyalkylene C 3 -C 12, dihydroxyalkylene C 4 -C 12, dialkylarylene C_ {8} -C_ {12}, - (R 1 O) x R 1 -, - (R 1 O) x R 5 (OR 1) x -, - (CH 2 CH (OR 2) CH 2 O) z - (R 1 O) and R 1 (OCH 2 CH (OR 2) CH 2) w -,  -C (O) (R 4) r C (O) -, -CH 2 CH (OR 2) CH 2 -, and mixtures of the themselves; wherein R 1 is alkylene C 2 -C 6 and mixtures thereof; R2 is hydrogen, - (R 1 O) x B, and mixtures thereof; R 3 is C 1 -C 18 alkyl, arylalkyl C 7 -C 12, alkyl substituted aryl C 7 -C 12, aryl C 6 -C 12, and mixtures thereof; R 4 it is C 1 -C 12 alkylene, alkenylene C 4 -C 12, arylalkylene C 8 -C 12, arylene 6 C 8 -C 10, and mixtures thereof; R 5 is C 1 -C 12 alkylene, C 3 -C 12 hydroxyalkylene, C4-C12 dihydroxyalkylene, dialkylarylene  C_ {8} -C_ {12}, -C (O) -, -C (O) NHR 6 NHC (O) -, -R 1 (OR 1) -, -C (O) (R 4) r C (O) -, -CH 2 CH (OH) CH 2 -, -CH 2 CH (OH) CH 2 O (R 1 O) and R 1 -OCH 2 CH (OH) CH 2 -, and mixtures thereof; R 6 is alkylene C 2 -C 12 or arylene C 6 -C 12; E units are selected from group consisting of hydrogen, alkyl C 1 -C 22, alkenyl C 3 -C 22, arylalkyl C 7 -C 22, hydroxyalkyl C 2 -C 22, - (CH 2) p CO 2 M, - (CH 2) q SO 3 M, -CH (CH 2 CO 2 M) CO 2 M, - (CH 2) p PO 3 M, - (R 1 O) x B, -C (O) R3, and mixtures thereof; as long as when any unit E of a nitrogen is a hydrogen, said Nitrogen is also not an N-oxide; B is hydrogen, alkyl C 1 -C 6, - (CH 2) q SO 3 M, - (CH 2) p CO 2 M, - (CH 2) q (CHSO 3 M) CH 2 SO 3 M, - (CH 2) q (CHSO 2 M) -CH 2 SO 3 M, - (CH 2) p PO 3 M, -PO 3 M, and mixtures of the themselves; M is hydrogen or a water-soluble cation in quantity enough to satisfy the load balance; X is an anion water soluble;

m has the value from 4 to 400; n has the value from 0 to 200; m + n has the value of al minus 5; p has the value from 1 to 6, q has the value from 0 to 6; r it has the value 0 or 1; w has the value 0 or 1; x has the value of 1 to 100; and has the value from 0 to 100; z has the value 0 or 1.

5. A laundry detergent composition according to any of the preceding claims, wherein the Dirt release polymer for cotton fabrics polyethyleneimine type is selected from polyethyleneimine 1800E7 and its derivatives of amine oxide, polyethyleneimine 1200E7 and its oxidized and / or quaternized derivatives, polyethyleneimine 600E20 and / or mixtures thereof. 6. A laundry detergent composition according to any of the preceding claims, which also comprises a surfactant, preferably a non-surfactant ionic. 7. A laundry detergent composition according to claim 6, wherein the nonionic surfactant is a non-ionic alkylethoxylated surfactant with a length of chain from C8 to C20, preferably from C12 to C16, and a degree of ethoxylation from 2 to 9, preferably from 3 to 7. 8. A laundry detergent composition according to claim 6, wherein the nonionic surfactant is a alkylmethyl glucamide surfactant with a length of C8 to C20 alkyl chain, preferably C12 to C18. 9. A laundry detergent composition according to any of the preceding claims, which also comprises a detergent builder additive, preferably a detergent builder additive selected from zeolite, sodium tripolyphosphate, laminar silicate and / or mixtures of same. 10. A laundry detergent composition according to any of the preceding claims, which also comprises a polymer for dirt release conventional, preferably a polyester with protected ends with anion, polypropylene diethoxylated terephthalate and / or mixtures of same. 11. A method of cleaning a tissue with a laundry detergent composition according to any of the previous claims.