BRPI0617599A2 - enzyme-catalyzed deacylation process of chlorinated sugar derivatives - Google Patents
enzyme-catalyzed deacylation process of chlorinated sugar derivatives Download PDFInfo
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- BRPI0617599A2 BRPI0617599A2 BRPI0617599-6A BRPI0617599A BRPI0617599A2 BR PI0617599 A2 BRPI0617599 A2 BR PI0617599A2 BR PI0617599 A BRPI0617599 A BR PI0617599A BR PI0617599 A2 BRPI0617599 A2 BR PI0617599A2
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- Prior art keywords
- sucrose
- chlorinated
- enzyme
- protected
- reaction
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Links
- 238000000034 method Methods 0.000 title claims abstract description 38
- 230000020176 deacylation Effects 0.000 title abstract description 24
- 238000005947 deacylation reaction Methods 0.000 title abstract description 24
- 102000004190 Enzymes Human genes 0.000 claims abstract description 22
- 108090000790 Enzymes Proteins 0.000 claims abstract description 22
- 238000005660 chlorination reaction Methods 0.000 claims abstract description 14
- 102000004882 Lipase Human genes 0.000 claims abstract description 11
- 108090001060 Lipase Proteins 0.000 claims abstract description 11
- 238000004519 manufacturing process Methods 0.000 claims abstract description 9
- 108091005804 Peptidases Proteins 0.000 claims abstract description 7
- 102100037486 Reverse transcriptase/ribonuclease H Human genes 0.000 claims abstract 4
- ZMXDDKWLCZADIW-UHFFFAOYSA-N Vilsmeier-Haack reagent Natural products CN(C)C=O ZMXDDKWLCZADIW-UHFFFAOYSA-N 0.000 claims description 55
- 238000006243 chemical reaction Methods 0.000 claims description 29
- 229930006000 Sucrose Natural products 0.000 claims description 16
- 239000005720 sucrose Substances 0.000 claims description 16
- CZMRCDWAGMRECN-UGDNZRGBSA-N Sucrose Chemical compound O[C@H]1[C@H](O)[C@@H](CO)O[C@@]1(CO)O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 CZMRCDWAGMRECN-UGDNZRGBSA-N 0.000 claims description 14
- 150000001875 compounds Chemical class 0.000 claims description 8
- 239000003153 chemical reaction reagent Substances 0.000 claims description 7
- 150000003445 sucroses Chemical class 0.000 claims description 7
- -1 chlorinated sucrose compound Chemical class 0.000 claims description 6
- 108010093096 Immobilized Enzymes Proteins 0.000 claims description 5
- 238000013019 agitation Methods 0.000 claims description 2
- UBOXGVDOUJQMTN-UHFFFAOYSA-N 1,1,2-trichloroethane Chemical compound ClCC(Cl)Cl UBOXGVDOUJQMTN-UHFFFAOYSA-N 0.000 claims 2
- XTEGARKTQYYJKE-UHFFFAOYSA-M Chlorate Chemical compound [O-]Cl(=O)=O XTEGARKTQYYJKE-UHFFFAOYSA-M 0.000 claims 2
- 125000006239 protecting group Chemical group 0.000 claims 2
- YBBRCQOCSYXUOC-UHFFFAOYSA-N sulfuryl dichloride Chemical compound ClS(Cl)(=O)=O YBBRCQOCSYXUOC-UHFFFAOYSA-N 0.000 claims 2
- RIOQSEWOXXDEQQ-UHFFFAOYSA-N triphenylphosphine Chemical compound C1=CC=CC=C1P(C=1C=CC=CC=1)C1=CC=CC=C1 RIOQSEWOXXDEQQ-UHFFFAOYSA-N 0.000 claims 2
- VEXZGXHMUGYJMC-UHFFFAOYSA-M Chloride anion Chemical compound [Cl-] VEXZGXHMUGYJMC-UHFFFAOYSA-M 0.000 claims 1
- AFHCRQREQZIDSI-OVUASUNJSA-N [(2r,3s,4s,5r,6r)-6-[(2s,3s,4s,5r)-3,4-dihydroxy-2,5-bis(hydroxymethyl)oxolan-2-yl]oxy-3,4,5-trihydroxyoxan-2-yl]methyl benzoate Chemical compound O[C@H]1[C@H](O)[C@@H](CO)O[C@@]1(CO)O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](COC(=O)C=2C=CC=CC=2)O1 AFHCRQREQZIDSI-OVUASUNJSA-N 0.000 claims 1
- 125000000217 alkyl group Chemical group 0.000 claims 1
- 125000001495 ethyl group Chemical group [H]C([H])([H])C([H])([H])* 0.000 claims 1
- 229940070765 laurate Drugs 0.000 claims 1
- 125000002496 methyl group Chemical group [H]C([H])([H])* 0.000 claims 1
- AFHCRQREQZIDSI-UHFFFAOYSA-N sucrose-6-benzoate Natural products OC1C(O)C(CO)OC1(CO)OC1C(O)C(O)C(O)C(COC(=O)C=2C=CC=CC=2)O1 AFHCRQREQZIDSI-UHFFFAOYSA-N 0.000 claims 1
- 239000011541 reaction mixture Substances 0.000 abstract description 9
- 238000006386 neutralization reaction Methods 0.000 abstract description 4
- 238000010979 pH adjustment Methods 0.000 abstract 1
- 229940088598 enzyme Drugs 0.000 description 18
- 230000002255 enzymatic effect Effects 0.000 description 12
- 238000004809 thin layer chromatography Methods 0.000 description 10
- 230000006196 deacetylation Effects 0.000 description 7
- 238000003381 deacetylation reaction Methods 0.000 description 7
- 238000002955 isolation Methods 0.000 description 6
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 6
- 230000002411 adverse Effects 0.000 description 4
- 230000015572 biosynthetic process Effects 0.000 description 4
- 230000006378 damage Effects 0.000 description 4
- 238000003756 stirring Methods 0.000 description 4
- 150000003511 tertiary amides Chemical class 0.000 description 4
- 229920003155 Eudragit® RL 100 Polymers 0.000 description 3
- 102000035195 Peptidases Human genes 0.000 description 3
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 description 3
- 230000008034 disappearance Effects 0.000 description 3
- 238000011084 recovery Methods 0.000 description 3
- 239000002904 solvent Substances 0.000 description 3
- CSCPPACGZOOCGX-UHFFFAOYSA-N Acetone Chemical compound CC(C)=O CSCPPACGZOOCGX-UHFFFAOYSA-N 0.000 description 2
- OYPRJOBELJOOCE-UHFFFAOYSA-N Calcium Chemical compound [Ca] OYPRJOBELJOOCE-UHFFFAOYSA-N 0.000 description 2
- DGAQECJNVWCQMB-PUAWFVPOSA-M Ilexoside XXIX Chemical compound C[C@@H]1CC[C@@]2(CC[C@@]3(C(=CC[C@H]4[C@]3(CC[C@@H]5[C@@]4(CC[C@@H](C5(C)C)OS(=O)(=O)[O-])C)C)[C@@H]2[C@]1(C)O)C)C(=O)O[C@H]6[C@@H]([C@H]([C@@H]([C@H](O6)CO)O)O)O.[Na+] DGAQECJNVWCQMB-PUAWFVPOSA-M 0.000 description 2
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 description 2
- MCMNRKCIXSYSNV-UHFFFAOYSA-N Zirconium dioxide Chemical compound O=[Zr]=O MCMNRKCIXSYSNV-UHFFFAOYSA-N 0.000 description 2
- 239000002253 acid Substances 0.000 description 2
- 239000003513 alkali Substances 0.000 description 2
- 239000002585 base Substances 0.000 description 2
- 239000011575 calcium Substances 0.000 description 2
- 229910052791 calcium Inorganic materials 0.000 description 2
- AXCZMVOFGPJBDE-UHFFFAOYSA-L calcium dihydroxide Chemical compound [OH-].[OH-].[Ca+2] AXCZMVOFGPJBDE-UHFFFAOYSA-L 0.000 description 2
- 239000000920 calcium hydroxide Substances 0.000 description 2
- 229910001861 calcium hydroxide Inorganic materials 0.000 description 2
- 239000012320 chlorinating reagent Substances 0.000 description 2
- 238000006911 enzymatic reaction Methods 0.000 description 2
- 238000002474 experimental method Methods 0.000 description 2
- 150000004679 hydroxides Chemical class 0.000 description 2
- 230000000813 microbial effect Effects 0.000 description 2
- 239000003960 organic solvent Substances 0.000 description 2
- CTSLXHKWHWQRSH-UHFFFAOYSA-N oxalyl chloride Chemical compound ClC(=O)C(Cl)=O CTSLXHKWHWQRSH-UHFFFAOYSA-N 0.000 description 2
- XHXFXVLFKHQFAL-UHFFFAOYSA-N phosphoryl trichloride Chemical compound ClP(Cl)(Cl)=O XHXFXVLFKHQFAL-UHFFFAOYSA-N 0.000 description 2
- 239000011148 porous material Substances 0.000 description 2
- 238000002360 preparation method Methods 0.000 description 2
- 239000000047 product Substances 0.000 description 2
- 229940024999 proteolytic enzymes for treatment of wounds and ulcers Drugs 0.000 description 2
- 239000002002 slurry Substances 0.000 description 2
- 239000011734 sodium Substances 0.000 description 2
- 229910052708 sodium Inorganic materials 0.000 description 2
- QTBSBXVTEAMEQO-UHFFFAOYSA-M Acetate Chemical compound CC([O-])=O QTBSBXVTEAMEQO-UHFFFAOYSA-M 0.000 description 1
- 229920001817 Agar Polymers 0.000 description 1
- 240000006439 Aspergillus oryzae Species 0.000 description 1
- 235000002247 Aspergillus oryzae Nutrition 0.000 description 1
- 241000193830 Bacillus <bacterium> Species 0.000 description 1
- 241000222120 Candida <Saccharomycetales> Species 0.000 description 1
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 description 1
- 229920002101 Chitin Polymers 0.000 description 1
- 229920001661 Chitosan Polymers 0.000 description 1
- KZBUYRJDOAKODT-UHFFFAOYSA-N Chlorine Chemical compound ClCl KZBUYRJDOAKODT-UHFFFAOYSA-N 0.000 description 1
- 229920002307 Dextran Polymers 0.000 description 1
- 241000222175 Diutina rugosa Species 0.000 description 1
- 241000196324 Embryophyta Species 0.000 description 1
- 239000004367 Lipase Substances 0.000 description 1
- 241001465754 Metazoa Species 0.000 description 1
- 239000004677 Nylon Substances 0.000 description 1
- 240000000064 Penicillium roqueforti Species 0.000 description 1
- 235000002233 Penicillium roqueforti Nutrition 0.000 description 1
- 239000004793 Polystyrene Substances 0.000 description 1
- ZLMJMSJWJFRBEC-UHFFFAOYSA-N Potassium Chemical compound [K] ZLMJMSJWJFRBEC-UHFFFAOYSA-N 0.000 description 1
- 101710091635 Probable diacyglycerol O-acyltransferase tgs1 Proteins 0.000 description 1
- 241000589517 Pseudomonas aeruginosa Species 0.000 description 1
- 241000235402 Rhizomucor Species 0.000 description 1
- 241000235545 Rhizopus niveus Species 0.000 description 1
- 229920002472 Starch Polymers 0.000 description 1
- 102000005158 Subtilisins Human genes 0.000 description 1
- 108010056079 Subtilisins Proteins 0.000 description 1
- 102100039146 Trimethylguanosine synthase Human genes 0.000 description 1
- 238000005917 acylation reaction Methods 0.000 description 1
- 230000006978 adaptation Effects 0.000 description 1
- 238000001042 affinity chromatography Methods 0.000 description 1
- 239000008272 agar Substances 0.000 description 1
- PNEYBMLMFCGWSK-UHFFFAOYSA-N aluminium oxide Inorganic materials [O-2].[O-2].[O-2].[Al+3].[Al+3] PNEYBMLMFCGWSK-UHFFFAOYSA-N 0.000 description 1
- 238000004458 analytical method Methods 0.000 description 1
- 239000008122 artificial sweetener Substances 0.000 description 1
- 235000021311 artificial sweeteners Nutrition 0.000 description 1
- 230000001580 bacterial effect Effects 0.000 description 1
- 229910052799 carbon Inorganic materials 0.000 description 1
- 239000000969 carrier Substances 0.000 description 1
- 239000001913 cellulose Substances 0.000 description 1
- 229920002678 cellulose Polymers 0.000 description 1
- 239000000460 chlorine Substances 0.000 description 1
- 229910052801 chlorine Inorganic materials 0.000 description 1
- 230000009849 deactivation Effects 0.000 description 1
- 238000006298 dechlorination reaction Methods 0.000 description 1
- 238000000354 decomposition reaction Methods 0.000 description 1
- 238000007257 deesterification reaction Methods 0.000 description 1
- 239000008367 deionised water Substances 0.000 description 1
- 229910021641 deionized water Inorganic materials 0.000 description 1
- 238000009826 distribution Methods 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- 125000004185 ester group Chemical group 0.000 description 1
- 238000000605 extraction Methods 0.000 description 1
- 239000012065 filter cake Substances 0.000 description 1
- 230000002538 fungal effect Effects 0.000 description 1
- 239000011521 glass Substances 0.000 description 1
- 150000004676 glycans Chemical class 0.000 description 1
- 238000004128 high performance liquid chromatography Methods 0.000 description 1
- 125000002887 hydroxy group Chemical group [H]O* 0.000 description 1
- 238000011065 in-situ storage Methods 0.000 description 1
- 235000019421 lipase Nutrition 0.000 description 1
- OHZZTXYKLXZFSZ-UHFFFAOYSA-I manganese(3+) 5,10,15-tris(1-methylpyridin-1-ium-4-yl)-20-(1-methylpyridin-4-ylidene)porphyrin-22-ide pentachloride Chemical compound [Cl-].[Cl-].[Cl-].[Cl-].[Cl-].[Mn+3].C1=CN(C)C=CC1=C1C(C=C2)=NC2=C(C=2C=C[N+](C)=CC=2)C([N-]2)=CC=C2C(C=2C=C[N+](C)=CC=2)=C(C=C2)N=C2C(C=2C=C[N+](C)=CC=2)=C2N=C1C=C2 OHZZTXYKLXZFSZ-UHFFFAOYSA-I 0.000 description 1
- 238000002156 mixing Methods 0.000 description 1
- 239000000203 mixture Substances 0.000 description 1
- 229920001778 nylon Polymers 0.000 description 1
- 230000002572 peristaltic effect Effects 0.000 description 1
- UHZYTMXLRWXGPK-UHFFFAOYSA-N phosphorus pentachloride Chemical compound ClP(Cl)(Cl)(Cl)Cl UHZYTMXLRWXGPK-UHFFFAOYSA-N 0.000 description 1
- 229920002401 polyacrylamide Polymers 0.000 description 1
- 229920001282 polysaccharide Polymers 0.000 description 1
- 239000005017 polysaccharide Substances 0.000 description 1
- 229920002223 polystyrene Polymers 0.000 description 1
- 239000011591 potassium Substances 0.000 description 1
- 229910052700 potassium Inorganic materials 0.000 description 1
- 239000002243 precursor Substances 0.000 description 1
- 239000000377 silicon dioxide Substances 0.000 description 1
- 239000008107 starch Substances 0.000 description 1
- 235000019698 starch Nutrition 0.000 description 1
- 125000003142 tertiary amide group Chemical group 0.000 description 1
- 229910000166 zirconium phosphate Inorganic materials 0.000 description 1
- LEHFSLREWWMLPU-UHFFFAOYSA-B zirconium(4+);tetraphosphate Chemical compound [Zr+4].[Zr+4].[Zr+4].[O-]P([O-])([O-])=O.[O-]P([O-])([O-])=O.[O-]P([O-])([O-])=O.[O-]P([O-])([O-])=O LEHFSLREWWMLPU-UHFFFAOYSA-B 0.000 description 1
Classifications
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07H—SUGARS; DERIVATIVES THEREOF; NUCLEOSIDES; NUCLEOTIDES; NUCLEIC ACIDS
- C07H3/00—Compounds containing only hydrogen atoms and saccharide radicals having only carbon, hydrogen, and oxygen atoms
- C07H3/04—Disaccharides
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07H—SUGARS; DERIVATIVES THEREOF; NUCLEOSIDES; NUCLEOTIDES; NUCLEIC ACIDS
- C07H1/00—Processes for the preparation of sugar derivatives
- C07H1/06—Separation; Purification
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07H—SUGARS; DERIVATIVES THEREOF; NUCLEOSIDES; NUCLEOTIDES; NUCLEIC ACIDS
- C07H5/00—Compounds containing saccharide radicals in which the hetero bonds to oxygen have been replaced by the same number of hetero bonds to halogen, nitrogen, sulfur, selenium, or tellurium
- C07H5/02—Compounds containing saccharide radicals in which the hetero bonds to oxygen have been replaced by the same number of hetero bonds to halogen, nitrogen, sulfur, selenium, or tellurium to halogen
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- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Engineering & Computer Science (AREA)
- Health & Medical Sciences (AREA)
- Biochemistry (AREA)
- Biotechnology (AREA)
- General Health & Medical Sciences (AREA)
- Genetics & Genomics (AREA)
- Molecular Biology (AREA)
- Life Sciences & Earth Sciences (AREA)
- Preparation Of Compounds By Using Micro-Organisms (AREA)
- Immobilizing And Processing Of Enzymes And Microorganisms (AREA)
- Enzymes And Modification Thereof (AREA)
- Saccharide Compounds (AREA)
Abstract
PROCESSO DE DESACILAçãO CATALISADA POR ENZIMAS DE DERIVADOS CLORADOS DE AçúCAR. Que consiste em um processo para a produção de triclorogalactosacarose, no qual a desacilação de sacarose-6-éster é obtida ao submeter a mistura de reação, após a cloração, a neutralização e o ajuste do pH entre 6,5 e 7, à desacilação, por meio do uso de uma enzima lípase ou de uma enzima protease, de uma forma livre ou imobilizada.DISACILATION PROCESS CATALYED BY ENZYMES OF CHLORINATED SUGAR DERIVATIVES. It consists of a process for the production of trichlorogalactosaccharose, in which sucrose-6-ester deacylation is obtained by subjecting the reaction mixture, after chlorination, neutralization and pH adjustment between 6.5 and 7, to deacylation , through the use of a lipase enzyme or a protease enzyme, in a free or immobilized form.
Description
PROCESSO DE DESACILAÇÃO CATALISADA POR ENZIMAS DE DERIVADOSCLORADOS DE AÇÚCAR"DISACILATION PROCESS CATALIZED BY SUGAR CHLORINE ENZYME "
Campo TécnicoTechnical Field
Trata-se a presente invenção de um processo inovador euma estratégia original para a produção de 1,6-dicloro-1-6-dideoxi-beta-fructofuranosil-4-cloro-4-deoxi-galactopiranosídeo (TGS), que envolve adesacilação enzimática de TGS 6-O-protegido obtido após a reação de cloração.This invention is an innovative process and an original strategy for the production of 1,6-dichloro-1-6-dideoxy-beta-fructofuranosyl-4-chloro-4-deoxy-galactopyranoside (TGS) involving enzymatic adhesacylation. of 6-O-protected TGS obtained after the chlorination reaction.
Fundamentos da InvençãoBackground of the Invention
As estratégias dos métodos da técnica anterior deprodução de 4, 1', 6'-triclorogalactosacarose (TGS) geralmente envolvem acloração de sacarose-6-éster por meio do uso do reagente Vilsmeier-Haackderivado de sacarose-6-éster clorada, para a formação de 6-acetil-4,16'-triclorogalactosacarose, por meio do uso de diversos agentes de cloração, como,por exemplo, oxicloreto de fósforo, cloreto de oxalila, pentacloreto de fósforo, etc.,e uma amida terciária, como, por exemplo, dimetilformamida (DMF). Após a ditareação de cloração, a massa de reação é neutralizada até atingir o pH entre 7,0 e7,7, por meio do uso de hidróxidos alcalinos apropriados de cálcio, sódio, etc. OpH da massa neutralizada é então aumentado para 9,5 ou mais, paradesesterificar/desacetilar 6-acetil-4,1', 6'-triclorogalactosacarose para formar 4,1',6'-triclorogalactosacarose, por meio do uso de hidróxidos alcalinos de cálcio,sódio, potássio, etc. Esta desesterificação/desacilação alcalina envolve aexposição dos reagentes a um pH adverso, na faixa alcalina, o que leva àdestruição de uma quantidade significativa de dimetilformamida (DMF), que é umacara contribuição, afetando adversamente sua recuperação após a reação.Prior art method strategies for the production of 4 ', 1', 6'-trichlorogalactosaccharide (TGS) generally involve sucrose-6-ester chlorination using Vilsmeier-Haackderived 6-ester chlorinated reagent for the formation of 6-acetyl-4,16'-trichlorogalactosacrosis by the use of various chlorinating agents such as phosphorus oxychloride, oxalyl chloride, phosphorus pentachloride etc. and a tertiary amide such as dimethylformamide (DMF). After chlorination dictation, the reaction mass is neutralized to pH 7.0 to 7.7 by the use of appropriate alkaline hydroxides of calcium, sodium, etc. The neutralized mass IH is then increased to 9.5 or more to esterify / deacetylate 6-acetyl-4,1 ', 6'-trichlorogalactosaccharide to form 4,1', 6'-trichlorogalactosaccharide by the use of alkaline hydroxides of calcium, sodium, potassium, etc. This alkaline deesterification / deacylation involves the exposure of reagents to an adverse pH in the alkaline range, which leads to the destruction of a significant amount of dimethylformamide (DMF), which is a contribution, adversely affecting their recovery after reaction.
No processo da técnica anterior, a mistura de reaçãotambém fica exposta durante o processo de desacilação a temperaturasadversas, que levam à destruição do próprio produto 4, 1', 6'-triclorogalactosacarose (TGS).Conseqüentemente, existe a necessidade de apresentarum método de desacilação que não exponha a dimetilformamida (DMF) àdestruição. Um método foi desenvolvido para a obtenção da desacilaçãoenzimática, a um pH que não exponha a dimetilformamida (DMF) à destruição.In the prior art process, the reaction mixture is also exposed during the deacylation process to reverse temperatures, which lead to the destruction of the product itself 4, 1 ', 6'-trichlorogalactosaccharide (TGS). Consequently, there is a need to present a deacylation method which does not expose dimethylformamide (DMF) to destruction. A method has been developed for obtaining enzymatic deacylation at a pH that does not expose dimethylformamide (DMF) to destruction.
Descrição Detalhada da InvençãoDetailed Description of the Invention
A desacilação enzimática foi descrita por Palmer e outros,em 1995, na Patente Norte-Americana No. 5445951, para a preparação dederivados parcialmente acilados de sacarose, por meio da desacilação catalisadapor enzimas de ésteres de sacarose a partir de um éster de sacarose selecionadodo grupo que consiste em octa-acilato de sacarose, hepta-acilato de sacarose ehexa-acilato de sacarose, em um meio orgânico anidro, com uma enzima ou comuma combinação de enzimas, que possa catalisar a desacilação do dito éster desacarose, para produzir um derivado de sacarose parcialmente desacilado, tendogrupo(s) hidroxila livre na(s) posição/posições pré-selecionada(s), e recuperar oderivado de sacarose parcialmente desacilado resultante.Enzymatic deacylation was described by Palmer et al. In 1995 in U.S. Patent No. 5,445,951 for the preparation of partially acylated sucrose derivatives by enzymatic deactivation of sucrose esters from a sucrose ester selected from the group. which consists of sucrose octaacylate, sucrose heptaacylate and sucrose achexate, in an anhydrous organic medium, with an enzyme or a combination of enzymes, which can catalyze the deacylation of said disaccharide ester to produce a partially deacylated sucrose, free hydroxyl group (s) at the preselected position (s), and recovering the resultant partially deacylated sucrose derivative.
Não há conhecimento de outro relatório a respeito dadesacilação enzimática de um éster de sacarose ou de seusderivados/precursores.No other report is known about the enzymatic desacilation of a sucrose ester or its derivatives / precursors.
A presente invenção se refere à desacilação enzimáticade TGS 6-O-protegido obtido após a reação de cloração durante a preparação doadoçante artificial, TGS. As modalidades da mistura da reação de cloração quepode ser submetida ao processo descrito na presente invenção incluem, mas nãoficam limitadas a um fluxo de tratamento obtido após a mistura de sacarose-6-éster com um agente de cloração, conforme descrito por Mufti e outros, em 1983,na Patente Norte-Americana No. 4.380.476, por Walkup e outros, em 1990, naPatente Norte-Americana No. 4.980.463, por Jenner e outros, em 1982, naPatente Norte-Americana No. 4.362.869, por Tulley e outros, em 1989, na PatenteNorte-Americana No. 4.801.700, por Rathbone e outros, em 1989, na PatenteNorte-Americana No. 4.826.962, por Bornemann e outros, em 1992, na PatenteNorte-Americana No. 5.141.860, por Navia e outros, em 1996, na Patente Norte-Americana No. 5.498.709, por Simpson, em 1989, na Patente Norte-AmericanaNo. 4.889.928, por Navia1 em 1990, na Patente Norte-Americana No. 4.950.746,por Neiditeh e outros, em 1991, na Patente Norte-Amerieana No. 5.023.329, porWalkup e outros, em 1992, 5.089.608, por Dordiek e outros, em 1992, na PatenteNorte-Amerieana No. 5.128.248, por Khan e outros, em 1995, na Patente Norte-Amerieana No. 5.440.026, por Palmer e outros, em 1995, na Patente Norte-Amerieana No. 5.445.951, por Sankey e outros, em 1995, na Patente Norte-Amerieana No. 5.449.772, por Sankey e outros, em 1995, na Patente Norte-Amerieana No. 5.470.969, por Navia e outros, em 1996, na Patente Norte-Americana No. 5.498.709, por Navia e outros, em 1996 e na Patente Norte-Americana No. 5.530.106. A desacilação enzimática é realizada no fluxo detratamento obtido conforme mencionado acima, após a neutralização da massada reação clorada, após ou sem o isolamento intermediário do TGS 6-0-protegido. O solvente, a amida terciária presente na massa da reaçãoneutralizada, não se decompõe devido à reação enzimática e, portanto, resulta narecuperação melhorada do dito solvente.The present invention relates to the 6-O-protected enzymatic deacylation obtained after the chlorination reaction during the preparation of artificial sweetener, TGS. Embodiments of the chlorination reaction mixture which may be subjected to the process described in the present invention include, but are not limited to, a treatment flow obtained after mixing sucrose-6-ester with a chlorinating agent as described by Mufti et al. in 1983, U.S. Patent No. 4,380,476, by Walkup et al., in 1990, U.S. Patent No. 4,980,463, by Jenner et al., in 1982, U.S. Patent No. 4,362,869, by Tulley et al. in 1989 in U.S. Patent No. 4,801,700; Rathbone et al. in 1989 in U.S. Patent No. 4,826,962; Bornemann et al. in 1992 in U.S. Patent No. 4,801,700; No. 5,141,860, by Navia et al., In 1996, in US Patent No. 5,498,709, by Simpson, in 1989, in U.S. Patent No. 5,441,860. 4,889,928, by Navia1 in 1990, U.S. Patent No. 4,950,746, by Neiditeh et al., In 1991, U.S. Patent No. 5,023,329, by Talkup et al., 1992, 5,089,608, by Dordiek et al. in 1992 in North American Patent No. 5,128,248, by Khan et al. in 1995 in North American Patent No. 5,440,026 by Palmer et al. in 1995 in North American Patent No. 5,445,951 by Sankey et al. In 1995 in U.S. Patent No. 5,449,772 by Sankey et al. In 1995 in U.S. Patent No. 5,470,969 by Navia et al. 1996, U.S. Patent No. 5,498,709, by Navia et al., In 1996 and U.S. Patent No. 5,530,106. Enzymatic deacylation is performed in the flow-through treatment obtained as mentioned above, after neutralization of the massive chlorinated reaction, after or without intermediate isolation of the 6-0 protected TGS. The solvent, the tertiary amide present in the neutralized reaction mass, does not decompose due to the enzymatic reaction and thus results in improved recovery of said solvent.
Na presente invenção, a massa da reação clorada, após areação de cloração, é neutralizada com uma base apropriada. Quando o pH écontrolado durante a neutralização abaixo de 6,0, o complexo TGS formado aindatem o grupo protegido intacto na 6! posição. O desagrupamento da 6! posição érealizado com ou sem isolamento do dito composto. Outras várias referênciastambém apontam que o desagrupamento pode ser efetuado com ou sem amidaterciária, bem como outros solventes e condições aquosas.In the present invention, the chlorinated reaction mass after chlorination sandblast is neutralized with an appropriate base. When the pH is controlled during neutralization below 6.0, the TGS complex formed gives the intact protected group at 6%. position. The ungrouping of 6! position is performed with or without isolation of said compound. Other various references also point out that the disassembly can be effected with or without amidaterium as well as other solvents and aqueous conditions.
A presente invenção descreve a desacilação na 6aposição por meio do uso de um processo enzimático, no qual a enzima remove,seletivamente, o grupo protegido na presença ou na ausência da amida terciária,incluindo dimetilformamida (DMF), que é utilizada na reação de cloração.The present invention describes the deacylation on position by the use of an enzymatic process in which the enzyme selectively removes the protected group in the presence or absence of tertiary amide, including dimethylformamide (DMF), which is used in the chlorination reaction. .
O processo da presente invenção também funciona bempara a desacilação de modalidades que não são resultantes de uma reação decloração, como, por exemplo, uma solução simples de TGS-6-éster puro.A desacilação catalisada por enzima é bastanteconhecida e as enzimas proteolíticas e as enzimas lípases realizam as reaçõesde desacilação e acilação sob condições benignas de reação e é amplamentedivulgado por Soedjak HS1 Spradlin JE (1994). Biocatalysis 11: 241-248; TherisodM. Klibanov AM (1986) J. Am. Chem. Soe. 108: 5638- 5640; B. Cambou e A.M.Klibanov1 J. Am. Chem. SOC., 106, 2687(1984); Kirpal S Bisht, Pure & Appl.Chem., Vol. 68, No. 3, pp. 749-752, 1996; F.J. Ploul; M.A. Crucesl,Biotechnology Letters 21: 635-639, 1999. Na presente invenção, após aneutralização da massa de reação, o pH é ajustado a 6,5, por meio do uso de uma base apropriada. A enzima lípase é então lentamente adicionada à massade reação sob agitação, à temperatura ambiente. A quantidade de enzimaadicionada à massa de reação varia entre 10% e 40% em peso/volume,dependendo das condições da reação e da atividade enzimática. O teor de amidaterciária na massa de reação neutralizada varia entre aproximadamente 10% e 40%. A mistura de reação é agitada continuamente por um período de 10 a 60horas, de preferência, entre 16 e 20 horas. A conversão de TGS 6-O-protegidoem TGS é monitorada por cromatografia em camada delgada (TLC). Após acompleta desacilação, a mistura de reação é levada para o isolamento do TGS,por meio de cromatografia de afinidade. O TGS isolado é então cristalizado por meio do uso de métodos apropriados.The process of the present invention also works well for the deacylation of modalities that are not the result of a dechlorination reaction, such as a simple solution of pure TGS-6-ester. Enzyme-catalyzed deacylation is well known and proteolytic enzymes and Lipase enzymes perform deacylation and acylation reactions under benign reaction conditions and is widely disclosed by Soedjak HS1 Spradlin JE (1994). Biocatalysis 11: 241-248; TherisodM. Klibanov AM (1986) J. Am. Chem. Sound. 108: 5638-5640; B. Cambou and A.M. Klibanov J. J. Am. Chem. SOC., 106, 2687 (1984); Kirpal S. Bisht, Pure & Appl.Chem., Vol. 68, No. 3, pp. 749-752, 1996; F.J. Ploul; M.A. Crucesl, Biotechnology Letters 21: 635-639, 1999. In the present invention, after neutralization of the reaction mass, the pH is adjusted to 6.5 by use of an appropriate base. The lipase enzyme is then slowly added to the mass under stirring at room temperature. The amount of enzyme added to the reaction mass ranges from 10% to 40% by weight / volume, depending on reaction conditions and enzyme activity. The amidatertiary content in the neutralized reaction mass ranges from approximately 10% to 40%. The reaction mixture is continuously stirred for a period of 10 to 60 hours, preferably 16 to 20 hours. Conversion of 6-O-protected TGS to TGS is monitored by thin layer chromatography (TLC). After complete deacylation, the reaction mixture is taken to isolation of the TGS by affinity chromatography. The isolated TGS is then crystallized by the use of appropriate methods.
O uso de enzimas lípases ou enzimas proteolíticas, para adesacilação de TGS 6-O-protegido em TGS, pode ser na sua forma nativa ou naforma imobilizada. Quando a enzima imobilizada é utilizada, a enzima é filtradaapós o término da desacilação. Esta enzima recuperada também pode ser reutilizada. Além disso, a enzima imobilizada também pode ser empacotada emuma coluna e a massa de reação pode ser passada através da coluna e adesacilação in situ do TGS 6-O-protegido pode ser realizada. Estas enzimaspodem ser imobilizadas em ou sobre suportes poliméricos sintéticos que incluem,por exemplo, mas não ficam limitados a suportes a base de náilon, poliacrílico,poliestireno ou poliacrilamida; ou suportes orgânicos naturais ou semi-sintéticos,como, por exemplo, aqueles baseados em polissacarídeos, que incluem, porexemplo, mas não ficam limitados a celulose, amido, dextrano, agár-agár,quitosana, quitina, etc; ou suportes inorgânicos, como, por exemplo, aquelesbaseados em carbono, sílica, zircônia, alumina, fosfato de zircônio, etc.The use of lipase enzymes or proteolytic enzymes for adhesion of TGS 6-O-protected TGS may be in its native or immobilized form. When the immobilized enzyme is used, the enzyme is filtered after the deacylation is complete. This recovered enzyme can also be reused. In addition, the immobilized enzyme may also be packaged in one column and the reaction mass may be passed through the column and in situ adhesionation of the 6-O-protected TGS may be performed. These enzymes may be immobilized on or on synthetic polymeric supports which include, for example, but are not limited to nylon, polyacrylic, polystyrene or polyacrylamide based supports; or natural or semi-synthetic organic carriers, such as those based on polysaccharides, which include, for example, but are not limited to cellulose, starch, dextran, agar, chitosan, chitin, etc .; or inorganic supports, such as those based on carbon, silica, zirconia, alumina, zirconium phosphate, etc.
A fonte das enzimas lípases pode ser de origem animal,vegetal ou microbiana, de preferência, de origem microbiana ou bacteriana, como,por exemplo, Bacillus thermocatenulatusis, Pseudomonas aeruginosa, etc., deorigem fúngica, como, por exemplo, Penicillium Roquefortii, Asperigillus niger,Asperigillus oryzae, Rhizopus niveus, Candida rugosa, Rhizomucor miheii,Candida antartctica, etc.The source of the lipase enzymes may be of animal, plant or microbial origin, preferably of microbial or bacterial origin, such as Bacillus thermocatenulatusis, Pseudomonas aeruginosa, etc., fungal origin such as Penicillium Roquefortii, Asperigillus. niger, Asperigillus oryzae, Rhizopus niveus, Candida rugosa, Rhizomucor miheii, Candida antarttica, etc.
Durante o processo da presente invenção, o TGS produtonão fica exposto a quaisquer condições adversas de temperatura ou pH, tal comono caso dos processos convencionais de desacilação, por meio do uso de ácido eálcali. A perda de produto é mínima comparada com qualquer outra forma deprocesso de desacilação.During the process of the present invention, TGS producon is not exposed to any adverse temperature or pH conditions, such as conventional deacylation processes, by the use of alkali acid. Product loss is minimal compared to any other form of deacylation process.
Durante o processo da presente invenção, a amidaterciária não fica exposta a quaisquer condições adversas de temperatura ou pH,tal como no caso dos processos convencionais de desacilação, por meio do usode ácido e álcali. Conseqüentemente, de forma alguma ocorre a decomposiçãoda amida terciária. Portanto, o rendimento da recuperação da amida terciáriaaumenta significativamente.During the process of the present invention, amidatertiary is not exposed to any adverse temperature or pH conditions, as in the case of conventional deacylation processes, by the use of acid and alkali. Consequently, the decomposition of tertiary amide does not occur at all. Therefore, the yield of tertiary amide recovery increases significantly.
Abaixo, são descritos exemplos que ilustram ofuncionamento da presente invenção, sem limitar, de qualquer maneira, o âmbitoda mesma. Os reagentes, a proporção dos reagentes utilizados, a variedade dascondições de reação descritas, as enzimas utilizadas e algo do gênero sãoapresentados apenas a título de ilustração e o âmbito da presente invençãoengloba seus reagentes análogos e condições de reação análogas, bem comoreações de natureza genérica análoga. Em geral, qualquer alternativaequivalente, que seja evidente para os especialistas versados na técnica deprodução de sacarose clorada, será incluída no âmbito do presente relatóriodescritivo. Conseqüentemente, a citação de um acetato abrangerá qualquer grupoéster equivalente que possa desempenhar a mesma função no contexto dapresente invenção, e a utilização de uma enzima abrangerá qualquer alternativaque possa fornecer a ação ou a ação análoga da enzima aqui descrita sobcondições análogas de reação. Várias outras adaptações das modalidades serãofacilmente previsíveis por aqueles especialistas versados nesta técnica e quetambém estão incluídas no âmbito do presente relatório descritivo. Uma citaçãona forma singular pretende também incluir o seu plural, a menos que o contextonão permita isso, ou seja: a utilização da expressão "um solvente orgânico" paraextração abrange a utilização de um ou mais de um solvente orgânico, sejasucessivamente ou em combinação, como uma mistura.Listed below are examples illustrating the operation of the present invention, without limiting in any way the scope thereof. The reagents, the proportion of reagents used, the variety of reaction conditions described, the enzymes used and the like are given by way of illustration only and the scope of the present invention encompasses analogous reagents and analogous reaction conditions as well as analogous generic reactions. . In general, any equivalent alternative, which is apparent to those skilled in the art of chlorinated sucrose production, will be included within the scope of this specification. Accordingly, the citation of an acetate will encompass any equivalent ester group which may perform the same function in the context of the present invention, and the use of an enzyme will encompass any alternative that may provide the action or analogous action of the enzyme described herein under analogous reaction conditions. Several other adaptations of the embodiments will be readily foreseeable by those skilled in the art and also included within the scope of this specification. A quotation in the singular form is also intended to include its plural unless the context otherwise permits, that is: the use of the term "an organic solvent" for extraction includes the use of one or more of an organic solvent, either successively or in combination, as a mix.
EXEMPLO 1EXAMPLE 1
Cloração de sacarose-6-acetatoSucrose-6-acetate chlorination
Em um frasco de reação de 5 litros, 1250 ml dedimetilformamida (DMF) foram adicionados e resfriados a uma temperatura entre0°C e 5°C. Depois disso, foram adicionados, lentamente, 635g de pentacloreto defósforo (5,4 moles), sob agitação, mantendo-se a temperatura da massa dereação inferior a 30°C. A massa também foi resfriada a uma temperatura inferior aO0C e a sacarose-6-acetato em dimetilformamida (DMF) foi lentamenteadicionada, a uma temperatura entre 0°C e 5°C. Em seguida, a massa de reaçãofoi aquecida a uma temperatura de 80°C e mantida por um período de uma hora,e depois também foi aquecida a uma temperatura de 100°C e mantida por umperíodo de 6 horas e, finalmente, foi aquecida a uma temperatura entre 1100C e115°C e mantida por um período entre 2 e 3 horas. O progresso da reação foicontrolado por análise por cromatografia líquida de alta eficiência (HPLC).In a 5 liter reaction flask, 1250 ml of dimethylformamide (DMF) was added and cooled to a temperature between 0 ° C and 5 ° C. Thereafter, 635g of dephosphorus pentachloride (5.4 moles) was slowly added under stirring, maintaining the temperature of the stripping mass below 30 ° C. The mass was also cooled to below 0 ° C and sucrose-6-acetate in dimethylformamide (DMF) was slowly added at a temperature between 0 ° C and 5 ° C. Thereafter, the reaction mass was heated to a temperature of 80 ° C and maintained for a period of one hour, and then also heated to a temperature of 100 ° C and maintained for a period of 6 hours and finally was heated to a temperature between 1100 ° C and 115 ° C and maintained for a period of 2 to 3 hours. The progress of the reaction was controlled by high performance liquid chromatography (HPLC) analysis.
Depois disso, a mistura de reação foi resfriada a umatemperatura entre -5°C e -8°C e uma solução de hidróxido de sódio 20% foiadicionada lentamente, de modo a levar o pH da massa entre 5,5 e 6,5. Orendimento obtido por este método foi de 55,4% de teor de sacarose.EXEMPLO 2After that, the reaction mixture was cooled to -5 ° C to -8 ° C and a slowly added 20% sodium hydroxide solution to bring the pH of the mass between 5.5 and 6.5. The yield obtained by this method was 55.4% sucrose content.EXAMPLE 2
Desacetilação enzimática de 6-O-acetil-TGS porenzimas lípasesEnzymatic deacetylation of 6-O-acetyl-TGS porase enzymes
A massa de reação, 1,5 litro, contendo 15g de TGS 6-0-acetilado preparado conforme descrição no Exemplo 1, foi neutralizada por meiodo uso de uma pasta de hidróxido de cálcio 50% até atingir o pH 7,5. A massa dereação neutralizada foi diluída a 6 litros, por meio do uso de água. O teor dedimetilformamida (DMF) foi de 33% na massa neutralizada. Foram isolados 84gde enzima lípase de Aspergillus oryzae ATCC 26850; acrescentou-se NCIM 1212à mistura de reação sob agitação contínua, à temperatura ambiente. A reação foicontinuada durante várias horas e a formação de TGS e o desaparecimento deTGS 6-O-acetilado foram monitorados por cromatografia em camada delgada(TLC). Ao final de 42 horas, obteve-se a desacilação de até 98,4%.The 1.5 liter reaction mass containing 15g of 6-0-acetylated TGS prepared as described in Example 1 was neutralized by the use of a 50% calcium hydroxide slurry to pH 7.5. The neutralized derating mass was diluted to 6 liters using water. The dimethylformamide (DMF) content was 33% by mass neutralized. 84g of Aspergillus oryzae ATCC 26850 lipase enzyme were isolated; NCIM 1212 was added to the reaction mixture under continuous stirring at room temperature. The reaction was continued for several hours and TGS formation and disappearance of 6-O-acetylated TGS were monitored by thin layer chromatography (TLC). At the end of 42 hours, the deacylation of up to 98.4% was obtained.
Após a desacilação, a massa foi levada para o isolamentode TGS, por meio de métodos apropriados.After deacylation, the mass was taken to TGS isolation by appropriate methods.
EXEMPLO 3EXAMPLE 3
Desacetilação enzimática de 6-O-acetil-TGS porenzimas lípases imobilizadas em Eudragit RL100Enzymatic deacetylation of 6-O-acetyl-TGS pore enzymes immobilized on Eudragit RL100
Em um experimento, 2,5 litros da massa de reaçãocontendo 80g de TGS 6-O-acetilado foram neutralizados por meio do uso de umapasta de hidróxido de cálcio 50% até atingir o pH 7,5. A massa de reaçãoneutralizada foi diluída a 6 litros por meio do uso de água. O teor dedimetilformamida (DMF) foi de 33% na massa neutralizada. Foram adicionados àmistura de reação, 120g de enzimas lípases imobilizadas em Eudragit RL100 sobagitação contínua a uma temperatura entre 25°C e 30°C, que normalmente é atemperatura ambiente. A reação foi continuada durante várias horas e aformação de TGS e o desaparecimento de TGS 6-O-acetilado foram monitoradospor cromatografia em camada delgada (TLC). Ao final de 24 horas, obteve-se adesacilação de até 98,3%.In one experiment, 2.5 liters of the reaction mass containing 80g of 6-O-acetylated TGS were neutralized by using a 50% calcium hydroxide slurry to pH 7.5. The neutralized reaction mass was diluted to 6 liters using water. The dimethylformamide (DMF) content was 33% by mass neutralized. To the reaction mixture was added 120g of lipase enzymes immobilized on Eudragit RL100 under continuous agitation at a temperature between 25 ° C and 30 ° C, which is usually at room temperature. The reaction was continued for several hours and TGS formation and disappearance of 6-O-acetylated TGS were monitored by thin layer chromatography (TLC). After 24 hours, adhesacylation of up to 98.3% was obtained.
A massa foi então filtrada e levada para o isolamento deTGS. A enzima obtida na torta de filtro foi lavada com água e armazenada paraser reutilizada.The mass was then filtered and taken to isolation of TGS. The enzyme obtained from the filter cake was washed with water and stored for reuse.
EXEMPLO 4EXAMPLE 4
Desacetilação enzimática de 6-0-acetil-TGS porenzimas lípases imobilizadas em Eudragit RL 100 empacotadas em umacolunaEnzymatic deacetylation of 6-0-acetyl-TGS pore enzymes immobilized on Eudragit RL 100 packed in a column
Em um experimento, 12g de enzimas imobilizadas foramempacotados em uma coluna de vidro de 2 cm de diâmetro e 8 cm de altura. Aentrada da coluna foi conectada ao ponto de distribuição de uma bombaperistáltica e a saída foi conectada a um frasco contendo 500 ml de massaneutralizada, que continha 5 g de 6-O-acetil. A entrada da bomba peristálticatambém foi conectada à massa neutralizada. A massa neutralizada foi circulada auma taxa de fluxo de 5 ml/minuto, através do leito de lípase imobilizada, durante 6horas.In one experiment, 12g of immobilized enzymes were packed in a 2 cm diameter 8 cm high glass column. The inlet of the column was connected to the distribution point of a bombaperistaltic and the outlet was connected to a flask containing 500 ml of mass neutralized, which contained 5 g of 6-O-acetyl. The peristaltic pump inlet was also connected to the neutralized ground. The neutralized mass was circulated at a flow rate of 5 ml / min through the immobilized lipase bed for 6 hours.
A cromatografia em camada delgada (TLC) foi realizada acada hora para ver o grau de desacetilação ocorrendo no frasco. Após 6 horas,foi observada uma desacetilação superior a 98%.Thin layer chromatography (TLC) was performed every hour to see the degree of deacetylation occurring in the flask. After 6 hours, a deacetylation greater than 98% was observed.
Após o término da desacetilação de 6-O-acetil-TGS emTGS1 o leito da enzima imobilizada foi lavado com água deionizada e foiarmazenada em acetona 10% em água, até nova utilização.After the deacetylation of 6-O-acetyl-TGS in TGS1 was completed, the immobilized enzyme bed was washed with deionized water and stored in 10% acetone in water until further use.
EXEMPLO 5EXAMPLE 5
Desacetilação enzimática de 6-O-acetil-TGS pelaenzima alcalase, uma enzima proteolíticaFoi recolhido 1,0 L de massa neutralizada após cloração,contendo 10g de TGS 6-O-acetilado para a reação enzimática. A massa dereação neutralizada foi diluída a 3 litros por meio do uso de água. Foramadicionados 200 ml de Alcalase 2,4L, uma enzima comercialmente obtida junto àNovozymes, derivada de B. Iichenformisl à mistura de reação, sob agitaçãocontínua, a uma temperatura entre 25°C e 30°C. A reação foi continuada durantevárias horas e a formação de TGS e o desaparecimento de TGS 6-O-acetiladoforam monitorados por cromatografia em camada delgada (TLC). Ao final de 36horas, obteve-se a desacilação de até 96,4%. Após a desacilação, a massa foilevada para o isolamento de TGS, por meio de métodos apropriados.Enzymatic deacetylation of 6-O-acetyl-TGS by the enzyme alkalase, a proteolytic enzyme 1.0 L of neutralized mass was collected after chlorination containing 10 g of 6-O-acetylated TGS for the enzymatic reaction. The neutralized derating mass was diluted to 3 liters using water. 200 ml of Alcalase 2,4L, an enzyme commercially obtained from Novovomes, derived from B. Iichenformisl, was added to the reaction mixture under continuous stirring at 25 ° C to 30 ° C. The reaction was continued for several hours and TGS formation and disappearance of 6-O-acetylad TGS were monitored by thin layer chromatography (TLC). At the end of 36 hours, the deacylation of up to 96.4% was obtained. After deacylation, the mass was taken to isolation of TGS by appropriate methods.
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| IN1175MU2005 | 2005-09-22 | ||
| PCT/IN2006/000385 WO2007054973A2 (en) | 2005-09-22 | 2006-09-21 | Enzyme catalyzed de-acylation of chlorinated sugar derivatives |
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| KR (1) | KR20080056187A (en) |
| CN (1) | CN101282983A (en) |
| AU (1) | AU2006313335A1 (en) |
| BR (1) | BRPI0617599A2 (en) |
| CA (1) | CA2623246A1 (en) |
| EA (1) | EA200800654A1 (en) |
| IL (1) | IL190251A0 (en) |
| LV (1) | LV13762B (en) |
| NO (1) | NO20081899L (en) |
| WO (1) | WO2007054973A2 (en) |
| ZA (1) | ZA200802518B (en) |
Families Citing this family (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102181494A (en) * | 2011-03-21 | 2011-09-14 | 盐城捷康三氯蔗糖制造有限公司 | Synthesis of sucrose-6-fatty acid ester through selective catalysis of immobilized aspergillus oryzae lipase |
Family Cites Families (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| GB8822674D0 (en) * | 1988-09-27 | 1988-11-02 | Tate & Lyle Plc | Preparation of acylated sucrose derivatives |
| US5445951A (en) * | 1993-02-02 | 1995-08-29 | Mcneil-Ppc, Inc. | Regioselective enzymatic deacylation of sucrose esters in anhydrous organic media |
-
2006
- 2006-09-21 AU AU2006313335A patent/AU2006313335A1/en not_active Abandoned
- 2006-09-21 BR BRPI0617599-6A patent/BRPI0617599A2/en not_active Application Discontinuation
- 2006-09-21 CA CA002623246A patent/CA2623246A1/en not_active Abandoned
- 2006-09-21 WO PCT/IN2006/000385 patent/WO2007054973A2/en not_active Ceased
- 2006-09-21 KR KR1020087008005A patent/KR20080056187A/en not_active Withdrawn
- 2006-09-21 JP JP2008531892A patent/JP2009508519A/en active Pending
- 2006-09-21 CN CNA2006800346126A patent/CN101282983A/en active Pending
- 2006-09-21 EA EA200800654A patent/EA200800654A1/en unknown
- 2006-09-21 EP EP06842764A patent/EP1940858A4/en not_active Withdrawn
-
2008
- 2008-03-18 ZA ZA200802518A patent/ZA200802518B/en unknown
- 2008-03-18 IL IL190251A patent/IL190251A0/en unknown
- 2008-04-09 LV LVP-08-59A patent/LV13762B/en unknown
- 2008-04-22 NO NO20081899A patent/NO20081899L/en not_active Application Discontinuation
Also Published As
| Publication number | Publication date |
|---|---|
| EP1940858A4 (en) | 2009-06-17 |
| CN101282983A (en) | 2008-10-08 |
| KR20080056187A (en) | 2008-06-20 |
| ZA200802518B (en) | 2009-06-24 |
| WO2007054973A3 (en) | 2007-07-12 |
| WO2007054973B1 (en) | 2007-08-23 |
| EA200800654A1 (en) | 2009-02-27 |
| WO2007054973A2 (en) | 2007-05-18 |
| NO20081899L (en) | 2008-06-23 |
| AU2006313335A1 (en) | 2007-05-18 |
| IL190251A0 (en) | 2008-11-03 |
| LV13762B (en) | 2009-01-20 |
| JP2009508519A (en) | 2009-03-05 |
| EP1940858A2 (en) | 2008-07-09 |
| CA2623246A1 (en) | 2007-05-18 |
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