WO1998020139A2 - Phytase obtenue a partir de germes de soja - Google Patents

Phytase obtenue a partir de germes de soja Download PDF

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Publication number
WO1998020139A2
WO1998020139A2 PCT/EP1997/006076 EP9706076W WO9820139A2 WO 1998020139 A2 WO1998020139 A2 WO 1998020139A2 EP 9706076 W EP9706076 W EP 9706076W WO 9820139 A2 WO9820139 A2 WO 9820139A2
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WO
WIPO (PCT)
Prior art keywords
phytase
soybean
precipitate
subjecting
amino acid
Prior art date
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Ceased
Application number
PCT/EP1997/006076
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English (en)
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WO1998020139A3 (fr
Inventor
Andrew J. Morgan
Martin Hessing
Hetty E. Sleijster-Selis
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Danisco UK Ltd
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Finnfeeds International Ltd
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Publication date
Application filed by Finnfeeds International Ltd filed Critical Finnfeeds International Ltd
Priority to BR9712731-0A priority Critical patent/BR9712731A/pt
Priority to CA002269773A priority patent/CA2269773A1/fr
Priority to EP97948873A priority patent/EP0942993A2/fr
Priority to AU70021/98A priority patent/AU7002198A/en
Publication of WO1998020139A2 publication Critical patent/WO1998020139A2/fr
Publication of WO1998020139A3 publication Critical patent/WO1998020139A3/fr
Anticipated expiration legal-status Critical
Ceased legal-status Critical Current

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Classifications

    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Y—ENZYMES
    • C12Y301/00—Hydrolases acting on ester bonds (3.1)
    • C12Y301/03—Phosphoric monoester hydrolases (3.1.3)
    • C12Y301/03026—4-Phytase (3.1.3.26), i.e. 6-phytase
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23K—FODDER
    • A23K20/00—Accessory food factors for animal feeding-stuffs
    • A23K20/10—Organic substances
    • A23K20/189—Enzymes
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES, NOT OTHERWISE PROVIDED FOR; PREPARATION OR TREATMENT THEREOF
    • A23L11/00—Pulses, i.e. fruits of leguminous plants, for production of food; Products from legumes; Preparation or treatment thereof
    • A23L11/30—Removing undesirable substances, e.g. bitter substances
    • A23L11/33—Removing undesirable substances, e.g. bitter substances using enzymes; Enzymatic transformation of pulses or legumes
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES, NOT OTHERWISE PROVIDED FOR; PREPARATION OR TREATMENT THEREOF
    • A23L5/00—Preparation or treatment of foods or foodstuffs, in general; Food or foodstuffs obtained thereby; Materials therefor
    • A23L5/20—Removal of unwanted matter, e.g. deodorisation or detoxification
    • A23L5/25—Removal of unwanted matter, e.g. deodorisation or detoxification using enzymes
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
    • C12N15/09—Recombinant DNA-technology
    • C12N15/63—Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
    • C12N15/79—Vectors or expression systems specially adapted for eukaryotic hosts
    • C12N15/82—Vectors or expression systems specially adapted for eukaryotic hosts for plant cells, e.g. plant artificial chromosomes (PACs)
    • C12N15/8241—Phenotypically and genetically modified plants via recombinant DNA technology
    • C12N15/8242—Phenotypically and genetically modified plants via recombinant DNA technology with non-agronomic quality (output) traits, e.g. for industrial processing; Value added, non-agronomic traits
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
    • C12N15/09—Recombinant DNA-technology
    • C12N15/63—Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
    • C12N15/79—Vectors or expression systems specially adapted for eukaryotic hosts
    • C12N15/82—Vectors or expression systems specially adapted for eukaryotic hosts for plant cells, e.g. plant artificial chromosomes (PACs)
    • C12N15/8241—Phenotypically and genetically modified plants via recombinant DNA technology
    • C12N15/8242—Phenotypically and genetically modified plants via recombinant DNA technology with non-agronomic quality (output) traits, e.g. for industrial processing; Value added, non-agronomic traits
    • C12N15/8257—Phenotypically and genetically modified plants via recombinant DNA technology with non-agronomic quality (output) traits, e.g. for industrial processing; Value added, non-agronomic traits for the production of primary gene products, e.g. pharmaceutical products, interferon
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
    • C12N9/14—Hydrolases (3)
    • C12N9/16—Hydrolases (3) acting on ester bonds (3.1)

Definitions

  • the invention relates to a novel class of phytate-degrading enzymes, in particular phytases (myo-inositol hexaphosphate hydrolases) , which are endogenously present in soya flour, soybeans, germinated soybeans or fractions thereof, to a method for obtaining such enzymes as well as to the use of these enzymes in feed and food technology.
  • phytases myo-inositol hexaphosphate hydrolases
  • Phytases are enzymes which catalyse the hydrolysis of phytic acid (myo-inositol hexakis phosphate) to myo- inositol and inorganic phosphate.
  • Phosphorous is an essential element for growth in animals.
  • Animal diets contain large amounts of phosphorous, the major part being present as phytate (phytic acid) which is largely not available for monogastric animals. Since phosphorous is essential for animal metabolism and the inorganic phosphate of the phytate molecule is largely unavailable to such animals, this dietary source of phosphorous passes through the digestive tract without contributing to the nutrition of said animals and is excreted in substantial quantities in the manure. Consequently, the application of large amounts of manure to farmland for agriculture purposes leads to an accumulation of phosphorous in the soil and to environmental pollution.
  • Phytases are produced by micro-organisms like Aspergillus niger (Vollova et al . , 1994), Aspergillus ficuum (Ullah and Dischinger, 1995) , Aspergillus carbonarius (Al-Asheh and Duvnjak, 1994), Klebsiella aerogenes (Tambe et al . , 1994) and Bacillus subtilis (Shimizu, 1992) .
  • Phytases are also produced by plants . Sutardi and Buckle (1986) and Gibson and Ullah (1988) have partially purified and characterised a phytase from soybeans . Chang and Swimmer (1976) have characterised phytase of Phaseolus vulgaris beans. Phytases were also found in peas (Beal and Mehta, 1985), barley malt (Lee, 1990), wheat (Khare et al . , 1994) and rye (Fretzdorff and Weipert, 1986) .Germination of certain seeds has been found to increase phytase activity.
  • the common sources of industrially available phytases are the fermentation broths of micro-organisms.
  • phytases in the feed industry has become increasingly important due to their phosphorous releasing activity on phytic acid present in feedstuffs.
  • the use of phytase serves to increase the availability of bound phosphate and complexed multivalent cations to monogastric animals. This leads to an increased bioavalability of phytate phosphate, better utilisation of the available nutrients in the animal diets, lowered phosphorous content of the manure and, consequently, reduces the impact of livestock production on the environment.
  • an object of the present invention is to provide a novel class of soybean phytase which has specific characteristics with respect to pH optima and temperature stability, etc., different from most microbial enzymes, and are therefore of importance for particular industrial applications.
  • a further object of the invention is to provide a method for obtaining and purifying phytase of the invention from soybeans or fractions thereof, soya flour, germinated soybeans or parts thereof .
  • a further object of the present invention is to provide DNA molecules encoding the phytase of the present invention, prokaryotic or eukaryotic organism or host cell transformed with a DNA molecule encoding phytase of the invention and capable of expressing said phytase and a recombinant method for producing phytase from said prokaryotic or eukaryotic organisms or host cells.
  • a further object of the invention is to provide for the use of the phytases of the invention for foods and animal feedstuffs and for reducing the environmental impact of phosphorous from livestock production and reducing the multivalent metal ion binding antinutritional effect of phytate .
  • the object of the present invention is solved by making available a phytase which is obtainable by extracting soybeans or fractions thereof, soya flour, germinated soybeans or parts thereof using an aqueous solvent and selectively precipitating proteinaceous material from the extract obtained or from a fraction thereof, and optionally further fractionating the precipitate.
  • Subject matter of the phytase of the invention is a soybean phytase which has an optimal pH of about 5.0 when measured in a buffer comprising 0.0091 M sodium phytate in 50 M acetic acid/NaOH and 1 mM CaCl 2 at 50°C for 4 hrs .
  • a further preferred embodiment of the phytase of the invention is that said phytase has a specific activity of at least 6 ⁇ mol/min/g protein, preferably 21 ⁇ mol/min/g protein, and more preferably, 1.3 mmol/min/g protein when measured in a buffer comprising 0.0091 M sodium phytate in 50 mM acetic acid/NaOH and 1 mM CaCl 2 at 50°C for 4 hrs at pH 5.0.
  • a further embodiment of the phytase of the present invention is that said phytase has a pi of about 4.9.
  • a further embodiment of the phytase of the present invention is that said phytase has a molecular weight of between 30,000 and 100,000 Daltons, preferably about 75,000 Daltons .
  • the phytase of the invention comprises an amino acid sequence in the N- terminal portion of the protein having at least 85% homology to the amino acid sequence given in SEQ ID NO : 1 and/or SEQ ID NO : 3, and, more preferably, comprises the amino acid sequence given in SEQ ID NO: 1 in the N-terminal portion of the protein and/or the internal amino acid sequence as given in SEQ ID NO : 3.
  • Subject matter of the present invention is also a purified phytase obtainable by a method comprising the steps of: a) subjecting an extract of soybeans or fractions thereof, soya flour, germinated soybeans or parts thereof to 50- 70% ammonium sulfate precipitation to form a precipitate comprising said phytase; b) subjecting said precipitate to anion exchange chromatography and collecting fractions containing said phytase; and c) subjecting said precipitate to cation exchange chromatography and collecting fractions containing said phytase .
  • the present invention relates to phytases with one or more of the above characteristics.
  • the present invention also provides a DNA molecule and vector DNA molecule encoding a phytase according to the invention as well as prokaryotic or eukaryotic organism or host cell transformed with said DNA molecule and capable of expressing said phytase.
  • said prokaryotic host cell is selected from the group comprising E. coli, Bacillus sp . , Lactobacillus sp. and Lactococcus sp .
  • said eukaryotic organism or host cell is a fungus selected from the group comprising Aspergillus, Trichoderma, Penicillium, Mucor, Kluyveromyces and Saccharomyces or is a plant selected from the group comprising soybean, corn and rapeseed and seeds thereof.
  • a method of producing phytase of the present invention comprising the steps of: a) subjecting an extract of soybeans or fractions thereof, soya flour, germinated soybeans or parts thereof to 50- 70% ammonium precipitation to form a precipitate comprising said phytase; b) subjecting said precipitate to anion exchange chromatography and collecting fractions containing said phytase ; and c) subjecting said precipitate to cation exchange chromatography and collecting fractions containing said phytase .
  • said anion exchange chromatography is performed using a Source Q column and/or said cation exchange chromatography is performed using a Source S column.
  • phytase and organisms of the present invention preferably plants or seeds, for reducing the environmental impact of phosphorous from livestock production and reducing the divalent metal ion binding antinutritional effect of phytate .
  • Figure 1 Protein content and phytase activity in fractions collected by chromatography on a Source Q column as described in Example 3.
  • Figure 2 Protein content and phytase activity in fractions collected by chromatography on a Source S column as described in Example 4.
  • Figure 3 SDS-PAGE analysis of the isolated phytase of the invention after column chromatography on Source Q and subsequent column chromatography on Source S; lane 1: molecular weight marker; lane 2: purified phytase, reduced; lane 3: purified phytase, non-reduced.
  • Figure 4 Isoelectric focusing of the isolated phytase of the invention after column chromatography on Source Q and subsequent column chromatography on Source S ; lane 1: molecular weight marker; lane 2: purified phytase.
  • Figure 5 The pH profile of the phytase of the invention after column chromatography on Source Q and subsequent column chromatography on Source S .
  • Soybeans 500 g, cv. Williams 82, Illinois Foundation Seeds, Champaign, II, USA
  • Soybeans were sterilised by washing with 0.5 % (w/v) sodium hypochloride for 30 minutes. Subsequently the beans were washed five times (five minutes each time) with sterile water.
  • the sterilised soybeans were germinated in six glass covered dishes with filter paper on the bottom (sterilised with 70% ethanol) at 20°C for 2 to 8 days in the dark. After germination, the soybeans were frozen at -24°C, lyophilised and milled in a Retch mill at 1 mm.
  • the milled soybeans were extracted with 0.05 M acetic acid/NaOH, pH 5.0, containing 1 mM CaCl 2 and 0.1% Tween-20, stirred at 20°C for 1.5 hrs, the suspension was centrifuged at 5°C at 16,000 g for 40 min and the resulting floating lipid layer was removed. The supernatant was fractionated by ammonium sulphate precipitation.
  • the protein pellet fraction obtained after 50 to 70 % ammonium sulphate precipitation contains proteins with a specific phytase activity of 6 ⁇ mol/min/g protein.
  • the protein content was measured with the Bio-Rad Protein assay (Bio-Rad, Veenendaal , the Netherlands) .
  • 1 ml of Bio- Rad Protein assay reagent (1 part was diluted with four parts H 2 0) was mixed with 25 ⁇ l of sample. After a period of 5 minutes to one hour the absorbence was measured at 595 nm. Ovalbumin was used for calibration.
  • the phytase activity was measured according to Simon et al . (1990) .
  • the specific phytase activity is described by the amount of phosphate which is liberated from 0.0091 M sodium phytate by 1 g protein at 50 °C and pH 5 during one minute under the conditions of the assay.
  • the phytase activity determination was carried out in 96 well microtiter plates.
  • Example 3 50 ⁇ l of sample was incubated with 100 ⁇ l of 0.0091 M sodium phytate in 50 mM acetic acid/NaOH, pH 5.0, containing 1 mM CaCl 2 at 50°C for 4 hrs. The incubation was stopped by adding 100 ⁇ l of a stop/colour reagent, containing 2.5% (w/v) ammonium heptamolybdate, 0.25% (w/v) ammonia (25%), 0.059% (w/v) ammonium vanadate and 6 % (v/v) nitric acid (65%) . The absorbance was measured at 415 nm against the blank incubation. Potassium dihydrogen phosphate was used for calibration (0-5 mM) .
  • Example 3 Example 3 :
  • the protein fraction obtained after 50 to 70 % ammonium sulphate precipitation was fractionated by an anion exchange chromatography (Source Q, Pharmacia, Uppsala, Sweden) column of 100 ml. 100 ml of protein fraction was desalted to the start elution buffer A (20 mM Tris/HCl buffer, pH 8.0) which was carried out by dialysing against the start buffer A. Subsequently, the dialysed fraction was centrifuged at 20,000 g for 20 minutes and the supernatant was diluted to 500 ml with start elution buffer A and applied to the column.
  • start elution buffer A (20 mM Tris/HCl buffer, pH 8.0
  • the first main peak pooled after the anion exchange chromatography was separated by a cation exchange chromatography (Source S, Pharmacia, Uppsala, Sweden) column of 20 ml.
  • the fraction pooled was desalted by a P6 column (Bio-Rad) and applied to the column by a 150 ml superloop.
  • Protein was detected by an UV detector at A280 nm and collected in 5 ml fractions.
  • the results of the chromatography on Source S are shown in Figure 2.
  • the phytase activity was measured in the collected fractions as described in example 2.
  • the isolated elution buffer A (20 mM acetic acid/Na
  • soya phytase showed a specific activity of about 1.3 mmol/min/g of protein.
  • the purified fraction was analysed by SDS-PAGE (gradient 10-15%) and isoelectric focusing (IEF) with a range from pH 4 to 6.5, using the Phast system of Pharmacia according to the instructions of the manufacturer.
  • the proteins were stained by silver staining. The results are shown in Figure 3 and 4 respectively.
  • Figure 3 shows that the reduced and non-reduced fraction consist of one protein band and demonstrated an apparent molecular weight of about 75,000 Da.
  • Figure 4 shows also one protein band with an isoelectric point of 4.9.
  • the purified fraction was also run on a 15% SDS-PAGE and blotted to Immobilon-P membrane in methanol/glycine transfer buffer after electrophoresis .
  • the membrane was washed five times with distilled water, stained with Coomassie Brilliant Blue R250 in 25% (v/v) methanol and 8% (w/v) acetic acid, destained, and the main protein band was excised from the membrane and the N-terminal amino acid sequence was determined with an Applied Biosystems mode 477 A gas-phase sequencer connected on-line to a 120 A PTH Analyser.
  • the sequencing of the N-terminus of the phytase according to the invention revealed the following sequence: H I P S T L E G P F D P V T V P F D P A L R G V A V D L P E T as given in SEQ ID NO : 1.
  • An internal fragment of the phytase according to the invention analyzed in the corresponding manner has the following sequence: F A D E P G H X P D P L S T P D P as presented in in SEQ ID NO : 3.
  • the cDNA of the gene encoding the soybean phytase of the invention is cloned using a nucleic acid probe based on the amino acid sequence as given in SEQ ID No : 1, preferably using a mixture of oligonucleotides comprising the DNA sequence GARGGNCCNT TYGAYCCNGT of Seq ID NO : 2 , where N is A, T, G, C or inosine, R is A or G and Y is T or C, and subsequently expressed in E. coli and soybean using procedures known in the art and described in Sambrook, J. et al .

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Abstract

Cette invention a trait à une catégorie d'enzymes de dégradation du phytate présentes de manière endogène dans la farine de soja, le soja, les germes de soja, ou dans leurs fractions, à un procédé d'obtention de ces enzymes et à leur utilsation dans des applications d'alimentation et de compléments nutritifs.
PCT/EP1997/006076 1996-11-05 1997-11-04 Phytase obtenue a partir de germes de soja Ceased WO1998020139A2 (fr)

Priority Applications (4)

Application Number Priority Date Filing Date Title
BR9712731-0A BR9712731A (pt) 1996-11-05 1997-11-04 Fitase a partir de feijões de soja germinados
CA002269773A CA2269773A1 (fr) 1996-11-05 1997-11-04 Phytase obtenue a partir de germes de soja
EP97948873A EP0942993A2 (fr) 1996-11-05 1997-11-04 Phytase obtenue a partir de germes de soja
AU70021/98A AU7002198A (en) 1996-11-05 1997-11-04 Phytase from germinated soybeans

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
GB9623133.7 1996-11-05
GB9623133A GB2319030A (en) 1996-11-05 1996-11-05 Phytase extracted from soybean

Publications (2)

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WO1998020139A2 true WO1998020139A2 (fr) 1998-05-14
WO1998020139A3 WO1998020139A3 (fr) 1998-07-23

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EP (1) EP0942993A2 (fr)
AU (1) AU7002198A (fr)
BR (1) BR9712731A (fr)
CA (1) CA2269773A1 (fr)
GB (1) GB2319030A (fr)
PL (1) PL333097A1 (fr)
WO (1) WO1998020139A2 (fr)

Cited By (26)

* Cited by examiner, † Cited by third party
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NL1006478C2 (nl) * 1997-01-20 1999-03-23 Bae Hee Dong Bereiding van enzymprodukten en ruwe voedingsmaterielen onder toepassing van graanzaden
WO1999067398A3 (fr) * 1998-06-25 2000-04-20 Cornell Res Foundation Inc Surexpression des genes de phytases dans des systemes de levures
WO2001004147A3 (fr) * 1999-07-12 2001-08-23 Rebecca E Cahoon Homologues de phosphatase de polyphosphate d'inositol vegetal
US6303766B1 (en) 1999-05-14 2001-10-16 Virginia Tech Intellectual Properties, Inc. Soybean phytase and nucleic acid encoding the same
WO2002000890A1 (fr) * 2000-06-30 2002-01-03 Biogemma Acides nucleiques codant pour une phosphatase vegetale de type mipp a activite phytasique et applications
WO2001083763A3 (fr) * 2000-05-04 2002-02-28 Forskningsct Risoe Polypeptides a activite phytase
US6511699B1 (en) 1999-03-31 2003-01-28 Cornell Research Foundation, Inc. Enzymes with improved phytase activity
US6841370B1 (en) 1999-11-18 2005-01-11 Cornell Research Foundation, Inc. Site-directed mutagenesis of Escherichia coli phytase
WO2005005646A2 (fr) 2003-06-10 2005-01-20 Novozymes North America, Inc. Processus et compositions de fermentation
US7309505B2 (en) 2002-09-13 2007-12-18 Cornell Research Foundation, Inc. Using mutations to improve Aspergillus phytases
US7833743B2 (en) 2001-10-31 2010-11-16 Phytex, Llc Phytase-containing animal food and method
EP2295545A1 (fr) 2002-09-26 2011-03-16 Novozymes North America, Inc. Procédés et compositions de fermentation
US7919297B2 (en) 2006-02-21 2011-04-05 Cornell Research Foundation, Inc. Mutants of Aspergillus niger PhyA phytase and Aspergillus fumigatus phytase
US8540984B2 (en) 2006-08-03 2013-09-24 Cornell Research Foundation, Inc. Phytases with improved thermal stability
WO2015035914A1 (fr) 2013-09-11 2015-03-19 Novozymes A/S Procédés de production de produits de fermentation
WO2017112540A1 (fr) 2015-12-22 2017-06-29 Novozymes A/S Procédés de production de produits de fermentation
WO2019055455A1 (fr) 2017-09-15 2019-03-21 Novozymes A/S Mélanges d'enzymes et procédés pour améliorer la qualité nutritionnelle d'aliments pour animaux
WO2019083831A1 (fr) 2017-10-23 2019-05-02 Novozymes A/S Procédés pour la réduction d'acide lactique dans un système de fermentation de biocarburant
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WO2021126966A1 (fr) 2019-12-16 2021-06-24 Novozymes A/S Procédés de production de produits de fermentation
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US7713562B2 (en) 2003-09-04 2010-05-11 Rose Acre Farms, Inc. Animal feed and methods for reducing ammonia and phosphorus levels in manure
EP4638725A1 (fr) 2022-12-19 2025-10-29 Novozymes A/S Polypeptides de la famille 3 de gludice estérase (ce3) présentant une activité acétyl xylane estérase et polynucléotides codant pour ceux-ci
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WO1999067398A3 (fr) * 1998-06-25 2000-04-20 Cornell Res Foundation Inc Surexpression des genes de phytases dans des systemes de levures
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KR100870886B1 (ko) * 1998-06-25 2008-11-28 코넬 리서치 파운데이션 인코포레이티드 효모 시스템에서 피타아제 유전자의 과량발현
EP1688500A3 (fr) * 1998-06-25 2008-11-05 Cornell Research Foundation, Inc. Surexpression de la Phytase dans la levure
US6451572B1 (en) 1998-06-25 2002-09-17 Cornell Research Foundation, Inc. Overexpression of phytase genes in yeast systems
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EP1749887A3 (fr) * 2000-05-04 2007-02-28 Forskningscenter Riso Polypeptides à activité phytase
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WO2001083763A3 (fr) * 2000-05-04 2002-02-28 Forskningsct Risoe Polypeptides a activite phytase
WO2002000890A1 (fr) * 2000-06-30 2002-01-03 Biogemma Acides nucleiques codant pour une phosphatase vegetale de type mipp a activite phytasique et applications
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EP2295545A1 (fr) 2002-09-26 2011-03-16 Novozymes North America, Inc. Procédés et compositions de fermentation
WO2005005646A2 (fr) 2003-06-10 2005-01-20 Novozymes North America, Inc. Processus et compositions de fermentation
US7919297B2 (en) 2006-02-21 2011-04-05 Cornell Research Foundation, Inc. Mutants of Aspergillus niger PhyA phytase and Aspergillus fumigatus phytase
US8540984B2 (en) 2006-08-03 2013-09-24 Cornell Research Foundation, Inc. Phytases with improved thermal stability
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BR9712731A (pt) 1999-10-26
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AU7002198A (en) 1998-05-29
WO1998020139A3 (fr) 1998-07-23
EP0942993A2 (fr) 1999-09-22
GB9623133D0 (en) 1997-01-08
PL333097A1 (en) 1999-11-08

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