WO2000023598A1 - Aminoacylase et son utilisation pour la preparation de d-acides amines - Google Patents
Aminoacylase et son utilisation pour la preparation de d-acides amines Download PDFInfo
- Publication number
- WO2000023598A1 WO2000023598A1 PCT/GB1999/003458 GB9903458W WO0023598A1 WO 2000023598 A1 WO2000023598 A1 WO 2000023598A1 GB 9903458 W GB9903458 W GB 9903458W WO 0023598 A1 WO0023598 A1 WO 0023598A1
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- WO
- WIPO (PCT)
- Prior art keywords
- enzyme
- acetyl
- substrate
- concentration
- process according
- Prior art date
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Classifications
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P13/00—Preparation of nitrogen-containing organic compounds
- C12P13/04—Alpha- or beta- amino acids
-
- 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)
- C12N9/78—Hydrolases (3) acting on carbon to nitrogen bonds other than peptide bonds (3.5)
- C12N9/80—Hydrolases (3) acting on carbon to nitrogen bonds other than peptide bonds (3.5) acting on amide bonds in linear amides (3.5.1)
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P41/00—Processes using enzymes or microorganisms to separate optical isomers from a racemic mixture
- C12P41/006—Processes using enzymes or microorganisms to separate optical isomers from a racemic mixture by reactions involving C-N bonds, e.g. nitriles, amides, hydantoins, carbamates, lactames, transamination reactions, or keto group formation from racemic mixtures
- C12P41/007—Processes using enzymes or microorganisms to separate optical isomers from a racemic mixture by reactions involving C-N bonds, e.g. nitriles, amides, hydantoins, carbamates, lactames, transamination reactions, or keto group formation from racemic mixtures by reactions involving acyl derivatives of racemic amines
Definitions
- This invention relates to an enzyme having D-aminoacylase activity and to its use in the production of D-aminoacids, by resolving a racemic mixture of N-acyl aminoacids and deprotecting optically-enriched N-acyl aminoacids.
- D-Aminoacids are commercially important intermediates in the production of various pesticides, antibiotics and other pharmaceuticals.
- phenylglycine and p-hydroxyphenylglycine are used in the synthesis of semi-synthetic penicillins and cephalosporins.
- D-aminoacids There is also much demand for novel D-aminoacids as building blocks for new drug substances.
- D-Aminoacids may be accessed by physical separation, for example by crystallisation of salts, or by asymmetric chemocatalysis by way of hydrogenation of an enamide precursor.
- Chemocatalysis provides a general method ofbroad applicability, e.g. for unnatural aminoacids, but requires subsequent N-deacylation for which conventional chemical hydrolysis often results in partial racemisation of the product.
- biocatalytic methods also, for example by the hydrolysis of hydantoins using a D-specific hydantoinase.
- the resulting D-carbamoyl aminoacid still requires enzymic or chemical deprotection to the aminoacid.
- L-aminoacids by means of an L-specific aminoacylase-catalysed hydrolysis of the racemic N-acetylaminoacid is a technology that is well established. This uses the enzyme from Aspergillus oryzae and has been operated on a commercial basis at very large scale, to produce L-methionine, L-valine and L-phenylalanine. Such a large- scale technology does not exist for production ofD-aminoacids, although D-aminoacylase activity has been identified in several microbial strains of Pseudomonas, Streptomyces and Alcaligenes. See Sugie and Suzuki, Agric. Biol. Chem.
- This enzyme obtained from Alcaligenes xylosoxydans subsp. xylosoxydans ( Alcaligenes A-6 M ), NCUvffi 10771, does not hydrolyse N-acetyl-D-tryptophan. It is reported that the activity of D-aminoacylase is inhibited by 37% and 40% by D- phenylalanine and N-acetyl-D-alloisoleucine at a very low concentration of 2mM. This suggests that the enzyme is susceptible to severe product and substrate inhibition.
- US-A-5206162 discloses a D-aminoacylase obtained from Alcaligenes faecalis, CCRC 14817.
- EP-A-0896057 discloses a D-aminoacylase obtained from Amycolatopsis orientalis, IFO 12806. Summary of the Invention
- the present invention was made following a screen for D-aminoacylase activity performed on a collection of bacteria, and from this screen several were identified as having a D-aminoacylase. Five of these strains were used for genomic DNA preparation. It was then possible, by examining a known literature sequence, to design oligonucleotide primers, and use these in PCR experiments to generate a 1.4kb fragment possessing D- aminoacylase activity. The recombinant fragment was sub-cloned into pTrc99C expression vector. The recombinant plasmid carrying the D-amino acylase fragment was then transformed in to E. coli DH5 for over-expression.
- the volume efficiency is low, which increases the cost of recovering the product and reduces the economic viability of the process.
- the enzyme is effective at lOOg/l of substrate; even at 200g/l good activity was demonstrated. It is useful at high volume efficiency, of about lOOg/l, for the deprotection of several (D)-N- acetylaminoacids. This allows an economical process to be developed.
- an isolated enzyme according to the present invention is capable of hydrolysing N-acetyl-D-tryptophan at a substrate concentration of 10 g 1.
- it is capable of the desired activity at the given concentration, and also at higher concentrations.
- it unlike the enzymes disclosed in US- A-5206162 and in EP-A- 0896057, it exhibits the ability to convert (R)-N-acetyl-2-thienylalanine, and also to convert it faster than (R)-N-acetyl-4-chlorophenylalanine.
- the substrate used in the invention may be part of a mixture of the (L)- and (D)-N-acylaminoacids.
- the (D)-N-acylaminoacid may be enantiomerically enriched, e.g. essentially optically pure.
- the novel enzyme may be used to produce natural and unnatural aminoacids.
- One class of the latter is aryl/heteroaryl-substituted aminoacids.
- the enzyme may suffer from substrate inhibition.
- a high substrate concentration may merely lead to a low conversion to product, so that a volume efficient reaction is not possible.
- this effect can be overcome by the simple expedient of adding the substrate in several batches over the course of thebiotransformation, and, if kept at low concentration, a high product accumulation is possible.
- substrate hydrolysis is poor.
- the enzyme will hydrolyse 15 g/1 efficiently and, by making several additions of the substrate, it is possible to accumulate about 75 g/1 of D-2-naphthylalanine.
- the enzyme may be used in whole cell or isolated form. It may be immobilised, if desired, by methods known to those of ordinary skill in the art.
- the enzyme may be produced from the deposited organism (details given below). Alternatively, it may be produced by recombinant technology.
- DNA and amino-acid sequence provided herein, a person skilled in the art can readily construct fragments or mutations of the genes and enzymes disclosed herein. These fragments and mutations, which retain the activity of the exemplified enzyme, are within the scope of the present invention. Also, because of the redundancy of the genetic code, a variety of different DNA sequences can encode the amino-acid sequences disclosed herein. It is well within the skill of one of ordinary skill in the art to create these alternative DNA sequences encoding the same, or similar, enzymes. These DNA sequences are within the scope of the present invention. As used herein, reference to "essentially the same" sequence refers to sequences which have amino-acid substitutions, deletions, additions or insertions which do not materially affect activity. Fragments retaining activity are also included in this definition.
- genes of this invention can be isolated by known procedures and can be introduced into a wide variety of microbial hosts. Expression of the gene results, directly or indirectly, in the intracellular production and maintenance of the enzyme.
- the gene may be introduced via a suitable vector into a microbial host.
- a DNA construct may include the transcriptional and translational regulatory signals for expression of the gene, the gene under their regulatory control and a DNA sequence homologous with a sequence in the host organism, whereby integration will occur, and/or a replication system which is functional in the host, whereby integration or stable maintenance will occur.
- the construct can involve the transcriptional regulatory region, if any, and the promoter, where the regulatory region may be either 5' or 3* of the promoter, the ribosomal binding site, the initiation codon, the structural gene having an open reading frame in phase with the initiation codon, the stop codon(s), the polyadenylation signal sequence, if any, and the terminator region.
- This sequence as a double strand may be used by itself for transformation of a microorganism host, but will usually be included with a DNA sequence involving a marker.
- the gene can be introduced between the transcriptional/translational initiation and termination regions, so as to be under the regulatory control of the initiation region.
- This construct can be included in a plasmid, which could include at least one replication system, but may include more than one, where one replication system is employed for cloning during the development of the plasmid and the second replication system is necessary for functioning in the ultimate host.
- one or more markers may be present, as described above.
- the plasmid will desirably include a sequence homologous with the host genome.
- the transformants can be isolated in accordance with conventional ways, usually employing a selection technique, which allows for selection of the desired organism as against unmodified organisms or transferring organisms, when present. The transformants then can be tested for activity.
- Suitable host cells include prokaryotes and eukaryotes.
- An example is E. coli.
- Genomic DNA was prepared from 5 Alcaligenes strains held in the Chirotech culture collection; CMC3352, 3353, 2916, 3378, 3823. From these genomic preparations PCR was carried out to amplify the D-aminoacylase reported by Wakayama et al (1995), supra. Primers were synthesised according to the published sequence of the dan gene from Alcaligenes A-6. The 5' PCR primer in SEQ ID NO. 1 ; the 3' PCR primer is SEQ ID NO. 2.
- a 1.4 kb PCR fragment was amplified from strains CMC 3352 and 3353. These fragments were then cloned into the PCR cloning vector from Stratagene, pCR-script and transformed into E. coli. Resultant clones were analysed by restriction mapping to ascertain the presence of a 1.4kb acylase fragment. Clones harbouring this fragment were sequenced to verify that the putative acylase showed homology to the reported sequence. DNA sequence analysis show the majority of cloned fragments to include SEQ ID NO. 3. The deduced aminoacid sequence is given below as SEQ ID NO. 4.
- residues of the recombinant D-acylase differ from the published sequence as follows; Ser 2 to Ala; Gin 3 to Glu; Ala 14 to Val; ; Gly 126 to Arg; Gly 240 to Arg; Glu 242 to Lys.
- the recombinant fragment was sub-cloned into pTrc99C expression vector via the 5' Nc ⁇ l and 3' BamY ⁇ . engineered restriction sites.
- the recombinant plasmid carrying the D-amino acylase fragment was transformed into E. coli DH5 for over-expression.
- the recombinant cells E. coli strain CMC 4406, have been deposited at NCIMB, 23 St. Machar Drive, Aberdeen AB243RY, Scotland.
- the accession number is NCIMB 40965.
- the recombinant cells were grown by fermentation in a medium containing glucose, peptides and salts. The seed culture was inoculated from plates, and incubated overnight in TSB medium containing 0.1 g/1 ampicillin at 37 °C.
- the inoculum (5ml, OD 5.0) was grown in 1.51 of the following medium which contained (amounts in g.l "1 unless otherwise indicated): KHTO 8
- the reaction was monitored by chiral GC as follows: 0.5 ml of the reaction mixture was taken and acidified to pH 2.0 with cone. HC1. The aqueous was extracted with EtOAc which was dried (MgSO 4 ) and filtered and treated with OJml of TMS-diazomethane. The derivatised product was assayed by chiral GC (Chrompack Chirasil L-Nal, 25m, 20psi He, 60°C for 10 mins, 5°C/min to 200°C, holding for 10 minutes, FID detection). After 1 hour the ee of the substrate had decreased to 68%, after 2 hours it was 24% and after 22 hours was 7%.
- Table 1 reports D-acylase reactions using a range of unnatural (R)-N-Ac- phenylalanine and (R)-N-Ac-alanine derivatives, and (R)-N-Ac-4-fluorophenylglycine.
- coli CMC4406 containing recombinant D-acylase were immobilised on a reactive soluble polymer (RSP).
- the RSP was prepared by reaction of polyethyleneimine (0.8g) with aqueous 25% w/v glutaraldehyde (1.6ml), to a total volume of 20 ml H 2 O.
- the RSP was then mixed with 1 Og of cells resuspended in 20ml H 2 O. This was stirred vigorously for 30 minutes, after which the immobilised cells, having the consistency of foam rubber, were recovered by filtration.
- the final product (20g) had a specific activity of 20.55 U/g and the recovery of activity was 43% of the whole cells used in the immobilisation.
- 1 Unit of activity is defined as the hydrolysis of 1 ⁇ mol/min N-Ac- D-tryptophan to D-tryptophan measured at a substrate concentration of lOmM at 25 °C, pH7.5.
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- Chemical & Material Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Organic Chemistry (AREA)
- Zoology (AREA)
- Engineering & Computer Science (AREA)
- Wood Science & Technology (AREA)
- Health & Medical Sciences (AREA)
- Genetics & Genomics (AREA)
- Bioinformatics & Cheminformatics (AREA)
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- General Engineering & Computer Science (AREA)
- Biotechnology (AREA)
- General Health & Medical Sciences (AREA)
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- Medicinal Chemistry (AREA)
- Molecular Biology (AREA)
- Biomedical Technology (AREA)
- Chemical Kinetics & Catalysis (AREA)
- General Chemical & Material Sciences (AREA)
- Analytical Chemistry (AREA)
- Enzymes And Modification Thereof (AREA)
- Preparation Of Compounds By Using Micro-Organisms (AREA)
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Abstract
Priority Applications (5)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
KR1020017004943A KR20010075649A (ko) | 1998-10-20 | 1999-10-20 | 아미노아실라제 및 이를 사용한 디-아미노산의 제조방법 |
AU62227/99A AU6222799A (en) | 1998-10-20 | 1999-10-20 | Aminoacylase and its use in the production of d-aminoacids |
EP99949259A EP1121446A1 (fr) | 1998-10-20 | 1999-10-20 | Aminoacylase et son utilisation pour la preparation de d-acides amines |
CA002347079A CA2347079A1 (fr) | 1998-10-20 | 1999-10-20 | Aminoacylase et son utilisation pour la preparation de d-acides amines |
JP2000577305A JP2002527110A (ja) | 1998-10-20 | 1999-10-20 | アミノアシラーゼおよび、d−アミノ酸の製造におけるその使用 |
Applications Claiming Priority (4)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
GBGB9822947.9A GB9822947D0 (en) | 1998-10-20 | 1998-10-20 | Aminoacylase and its use in the production of d-aminoacids |
GBGB9907739.8A GB9907739D0 (en) | 1999-04-01 | 1999-04-01 | Aminoacylase and its use in the production of D-aminoacids |
GB9907739.8 | 1999-04-01 | ||
GB9822947.9 | 1999-04-01 |
Publications (1)
Publication Number | Publication Date |
---|---|
WO2000023598A1 true WO2000023598A1 (fr) | 2000-04-27 |
Family
ID=26314541
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
PCT/GB1999/003458 WO2000023598A1 (fr) | 1998-10-20 | 1999-10-20 | Aminoacylase et son utilisation pour la preparation de d-acides amines |
Country Status (6)
Country | Link |
---|---|
EP (1) | EP1121446A1 (fr) |
JP (1) | JP2002527110A (fr) |
KR (1) | KR20010075649A (fr) |
AU (1) | AU6222799A (fr) |
CA (1) | CA2347079A1 (fr) |
WO (1) | WO2000023598A1 (fr) |
Cited By (4)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
EP1120465A1 (fr) * | 2000-01-27 | 2001-08-01 | Daicel Chemical Industries, Ltd. | D-aminoacylase et gène codant pour celle-ci |
WO2002061077A1 (fr) * | 2001-02-01 | 2002-08-08 | Mitsui Chemicals, Inc. | Adn codant une nouvelle d-aminoacylase et procede pour produire un d-aminoacide au moyen de cet adn |
EP1435388A2 (fr) | 2002-12-24 | 2004-07-07 | Daicel Chemical Industries, Ltd. | Mutants de la D-aminoacylase d'Alcaligenes denitrificans pour une meilleure producion d'aminoacides-D |
CN108624577A (zh) * | 2017-03-22 | 2018-10-09 | 中国科学院天津工业生物技术研究所 | 用于催化n-乙酰-d-色氨酸水解生成d-色氨酸的新酶 |
Families Citing this family (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
KR20220096237A (ko) | 2020-12-30 | 2022-07-07 | 이종학 | 수산물 유통 서비스 시스템 |
Citations (8)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
DE2344060A1 (de) * | 1972-08-31 | 1974-03-07 | Ajinomoto Kk | Verfahren zur herstellung eines optisch aktiven enantiomorphen einer aminosaeure |
DE2419838A1 (de) * | 1973-04-24 | 1974-11-21 | Ajinomoto Kk | Verfahren zur herstellung einer optisch aktiven aminosaeure |
JPS62126976A (ja) * | 1985-11-26 | 1987-06-09 | Agency Of Ind Science & Technol | D−アミノアシラ−ゼの製造法 |
JPS645488A (en) * | 1987-06-29 | 1989-01-10 | Daicel Chem | Novel d-aminoacylase and production thereof |
EP0304021A2 (fr) * | 1987-08-21 | 1989-02-22 | Takeda Chemical Industries, Ltd. | Racémase d'un acide aminé acylé, préparation et utilisation |
JPH02234677A (ja) * | 1989-03-07 | 1990-09-17 | Dai Ichi Pure Chem Co Ltd | 酸性d―アミノ酸に作用するd―アミノアシラーゼ及びその製造法 |
US5206162A (en) * | 1991-10-17 | 1993-04-27 | National Science Council Of Republic Of China | Process for making D-aminoacylase |
EP0896057A2 (fr) * | 1997-07-31 | 1999-02-10 | DAICEL CHEMICAL INDUSTRIES, Ltd. | D-aminoacylase |
-
1999
- 1999-10-20 WO PCT/GB1999/003458 patent/WO2000023598A1/fr not_active Application Discontinuation
- 1999-10-20 CA CA002347079A patent/CA2347079A1/fr not_active Abandoned
- 1999-10-20 AU AU62227/99A patent/AU6222799A/en not_active Abandoned
- 1999-10-20 KR KR1020017004943A patent/KR20010075649A/ko not_active Application Discontinuation
- 1999-10-20 EP EP99949259A patent/EP1121446A1/fr not_active Withdrawn
- 1999-10-20 JP JP2000577305A patent/JP2002527110A/ja active Pending
Patent Citations (8)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
DE2344060A1 (de) * | 1972-08-31 | 1974-03-07 | Ajinomoto Kk | Verfahren zur herstellung eines optisch aktiven enantiomorphen einer aminosaeure |
DE2419838A1 (de) * | 1973-04-24 | 1974-11-21 | Ajinomoto Kk | Verfahren zur herstellung einer optisch aktiven aminosaeure |
JPS62126976A (ja) * | 1985-11-26 | 1987-06-09 | Agency Of Ind Science & Technol | D−アミノアシラ−ゼの製造法 |
JPS645488A (en) * | 1987-06-29 | 1989-01-10 | Daicel Chem | Novel d-aminoacylase and production thereof |
EP0304021A2 (fr) * | 1987-08-21 | 1989-02-22 | Takeda Chemical Industries, Ltd. | Racémase d'un acide aminé acylé, préparation et utilisation |
JPH02234677A (ja) * | 1989-03-07 | 1990-09-17 | Dai Ichi Pure Chem Co Ltd | 酸性d―アミノ酸に作用するd―アミノアシラーゼ及びその製造法 |
US5206162A (en) * | 1991-10-17 | 1993-04-27 | National Science Council Of Republic Of China | Process for making D-aminoacylase |
EP0896057A2 (fr) * | 1997-07-31 | 1999-02-10 | DAICEL CHEMICAL INDUSTRIES, Ltd. | D-aminoacylase |
Non-Patent Citations (7)
Title |
---|
M. MORIGUCHI ET AL.: "Production, purification and characterization of D-aminoacylase from Alcaligenes xylosoxydans subsp. xylosoxydans A-6", BIOSCIENCE, BIOTECHNOLOGY AND BIOCHEMISTRY, vol. 57, July 1993 (1993-07-01), pages 1149 - 1152, XP002105915 * |
M. SUGIE ET AL.: "Optical resolution of DL-amino acids with D-aminoacylase of Streptomyces.", AGRICULTURAL & BIOLOGICAL CHEMISTRY, vol. 44, no. 5, 1980, pages 1089 - 1095, XP002105917 * |
M. WAKAYAMA ET AL.: "Cloning and sequencing of a gene encoding D-aminoacylase from Alcaligenes xylosoxydans subsp. sylososydans A-6 and expression of the gene in Escherichia coli.", BIOSCIENCE, BIOTECHNOLOGY AND BIOCHEMISTRY, vol. 59, November 1995 (1995-11-01), pages 2115 - 2119, XP002105914 * |
M. WAKAYAMA ET AL.: "Overproduction of D-aminoacylase from Alcaligenes xylosoxydans subsp. xylosoxydans A-6 in Escherichia coli and its purification.", PROTEIN EXPRESSION AND PURIFICATION, vol. 7, 1996, pages 395 - 399, XP002105916 * |
PATENT ABSTRACTS OF JAPAN vol. 011, no. 353 (C - 457) 18 November 1987 (1987-11-18) * |
PATENT ABSTRACTS OF JAPAN vol. 013, no. 174 (C - 589) 25 April 1989 (1989-04-25) * |
PATENT ABSTRACTS OF JAPAN vol. 014, no. 548 (C - 0785) 5 December 1990 (1990-12-05) * |
Cited By (10)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
EP1120465A1 (fr) * | 2000-01-27 | 2001-08-01 | Daicel Chemical Industries, Ltd. | D-aminoacylase et gène codant pour celle-ci |
US6780619B2 (en) | 2000-01-27 | 2004-08-24 | Daicel Chemical Industries, Ltd. | D-aminoacylase and gene encoding the same |
US6887697B2 (en) | 2000-01-27 | 2005-05-03 | Daicel Chemical Industries, Ltd. | D-aminoacylase and gene encoding the same |
WO2002061077A1 (fr) * | 2001-02-01 | 2002-08-08 | Mitsui Chemicals, Inc. | Adn codant une nouvelle d-aminoacylase et procede pour produire un d-aminoacide au moyen de cet adn |
US6869788B2 (en) | 2001-02-01 | 2005-03-22 | Mitsui Chemicals, Inc. | DNA encoding novel D-aminoacylase and process for producing D-amino acid by using the same |
EP1435388A2 (fr) | 2002-12-24 | 2004-07-07 | Daicel Chemical Industries, Ltd. | Mutants de la D-aminoacylase d'Alcaligenes denitrificans pour une meilleure producion d'aminoacides-D |
EP1435388A3 (fr) * | 2002-12-24 | 2004-10-13 | Daicel Chemical Industries, Ltd. | Mutants de la D-aminoacylase d'Alcaligenes denitrificans pour une meilleure producion d'aminoacides-D |
US7282356B2 (en) | 2002-12-24 | 2007-10-16 | Daicel Chemical Industries, Ltd. | D-aminoacylase mutants |
CN108624577A (zh) * | 2017-03-22 | 2018-10-09 | 中国科学院天津工业生物技术研究所 | 用于催化n-乙酰-d-色氨酸水解生成d-色氨酸的新酶 |
CN108624577B (zh) * | 2017-03-22 | 2021-07-27 | 中国科学院天津工业生物技术研究所 | 用于催化n-乙酰-d-色氨酸水解生成d-色氨酸的新酶 |
Also Published As
Publication number | Publication date |
---|---|
KR20010075649A (ko) | 2001-08-09 |
EP1121446A1 (fr) | 2001-08-08 |
CA2347079A1 (fr) | 2000-04-27 |
JP2002527110A (ja) | 2002-08-27 |
AU6222799A (en) | 2000-05-08 |
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