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1. (WO2017020839) HETEROLOGOUS EXPRESSION AND USE OF D-ACETYLGLUCOSAMINE DEACETYLASE
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Pub. No.: WO/2017/020839 International Application No.: PCT/CN2016/093103
Publication Date: 09.02.2017 International Filing Date: 03.08.2016
Chapter 2 Demand Filed: 05.06.2017
IPC:
C12N 9/80 (2006.01) ,C12N 15/70 (2006.01) ,C12N 1/21 (2006.01) ,C12P 19/26 (2006.01) ,C12R 1/19 (2006.01)
C CHEMISTRY; METALLURGY
12
BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
N
MICRO-ORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICRO-ORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
9
Enzymes, e.g. ligases (6.); Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating, or purifying enzymes
14
Hydrolases (3.)
78
acting on carbon to nitrogen bonds other than peptide bonds (3.5)
80
acting on amide bonds in linear amides
C CHEMISTRY; METALLURGY
12
BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
N
MICRO-ORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICRO-ORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
15
Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
09
Recombinant DNA-technology
63
Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
70
Vectors or expression systems specially adapted for E. coli
C CHEMISTRY; METALLURGY
12
BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
N
MICRO-ORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICRO-ORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
1
Micro-organisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving micro-organisms or compositions thereof; Processes of preparing or isolating a composition containing a micro-organism; Culture media therefor
20
Bacteria; Culture media therefor
21
modified by introduction of foreign genetic material
C CHEMISTRY; METALLURGY
12
BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
P
FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
19
Preparation of compounds containing saccharide radicals
26
Preparation of nitrogen-containing carbohydrates
C CHEMISTRY; METALLURGY
12
BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
R
INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C-C12Q75
1
Micro-organisms
01
Bacteria or actinomycetales
185
Escherichia
19
Escherichia coli
Applicants:
南京农业大学 NANJING AGRICULTURAL UNIVERSITY [CN/CN]; 中国江苏省南京市 玄武区卫岗1号 No.1 Weigang, Xuanwu Nanjing, Jiangsu 210095, CN
Inventors:
弗戈⋅迈约瑟夫 VOGLMEIR, Josef; CN
刘丽 LIU, Li; CN
吕永梅 LV, Yongmei; CN
拉博德⋅佩德罗 LABORDA, Pedro; CN
Agent:
南京天华专利代理有限责任公司 NANJING TIANHUA PATENT AGENT CO., LTD.; 中国江苏省南京市 鼓楼区山西路67号世贸中心大厦A1座23层 Floor 23, Block A1, World Trade Center Building, No. 67 Shanxi Road, Gulou Nanjing, Jiangsu 210009, CN
Priority Data:
201510472229.X04.08.2015CN
Title (EN) HETEROLOGOUS EXPRESSION AND USE OF D-ACETYLGLUCOSAMINE DEACETYLASE
(FR) EXPRESSION HÉTÉROLOGUE ET UTILISATION DE LA D-ACÉTYLGLUCOSAMINE DÉSACÉTYLASE
(ZH) 一种D-乙酰氨基葡萄糖脱乙酰基酶的异源表达及其应用
Abstract:
(EN) Provided are a heterologous expression and use of a good quality D-acetylglucosamine deacetylase. The D-acetylglucosamine deacetylase has an amino acid sequence as shown in SEQNO.1. An analysis on an assay on enzyme activity and substrate specificity shows that the D-acetylglucosamine deacetylase has a high substrate specificity. Regarding substrates in the study, deacetylation is specifically performed on a D-acetylglucosamine (GlcNAc) and a β-linked D-GlcNAc to produce a D-glucosamine (GlcN) or a β-linked GlcN. A means of genetic engineering is used to clone and express a recombinant D-acetylglucosamine deacetylase, thereby realizing a high expression level of the D-acetylglucosamine deacetylase, and obtaining a good quality D-acetylglucosamine deacetylase.
(FR) L'invention concerne une expression hétérologue et une utilisation d'une D-acétylglucosamine désacétylase de bonne qualité. La D-acétylglucosamine désacétylase présente une séquence d'acides aminés telle que montrée dans la séquence SEQ NO 1. Une analyse sur un essai d'activité enzymatique et de spécificité du substrat montre que la D-acétylglucosamine désacétylase présente une spécificité élevée de substrat. En ce qui concerne les substrats dans l'étude, la désacétylation est spécifiquement réalisée sur une D-acétylglucosamine (GlcNAc) et une D-GlcNAc liée en β pour produire une D-glucosamine (GlcN) ou une GlcN liée en β. Un moyen de génie génétique est utilisé pour cloner et exprimer une D-acétylglucosamine désacétylase recombinante, ce qui permet de réaliser un niveau d'expression élevé de la D-acétylglucosamine désacétylase et d'obtenir une D-acétylglucosamine désacétylase de bonne qualité.
(ZH) 一种优质D-乙酰氨基葡萄糖脱乙酰基酶的异源表达及其应用,所述D-乙酰氨基葡萄糖脱乙酰基酶的氨基酸序列如SEQNO.1所示。通过酶活性测定和底物特异性的分析,所述D-乙酰氨基葡萄糖脱乙酰基酶底物特异性专一强,所研究底物中针对性的将D-乙酰氨基葡萄糖(GlcNAc)以及β键连接的D-乙酰氨基葡萄糖(GlcNAc)脱乙酰化生成D-氨基葡萄糖(GlcN)或者β键连接的氨基葡萄糖(GlcN)。利用基因工程手段克隆表达了重组D-乙酰氨基葡萄糖脱乙酰基酶,实现了D-乙酰氨基葡萄糖脱乙酰基酶的大量表达,获得了优质的D-乙酰氨基葡萄糖脱乙酰基酶。
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Designated States: AE, AG, AL, AM, AO, AT, AU, AZ, BA, BB, BG, BH, BN, BR, BW, BY, BZ, CA, CH, CL, CN, CO, CR, CU, CZ, DE, DK, DM, DO, DZ, EC, EE, EG, ES, FI, GB, GD, GE, GH, GM, GT, HN, HR, HU, ID, IL, IN, IR, IS, JP, KE, KG, KN, KP, KR, KZ, LA, LC, LK, LR, LS, LU, LY, MA, MD, ME, MG, MK, MN, MW, MX, MY, MZ, NA, NG, NI, NO, NZ, OM, PA, PE, PG, PH, PL, PT, QA, RO, RS, RU, RW, SA, SC, SD, SE, SG, SK, SL, SM, ST, SV, SY, TH, TJ, TM, TN, TR, TT, TZ, UA, UG, US, UZ, VC, VN, ZA, ZM, ZW
African Regional Intellectual Property Organization (ARIPO) (BW, GH, GM, KE, LR, LS, MW, MZ, NA, RW, SD, SL, ST, SZ, TZ, UG, ZM, ZW)
Eurasian Patent Office (AM, AZ, BY, KG, KZ, RU, TJ, TM)
European Patent Office (EPO) (AL, AT, BE, BG, CH, CY, CZ, DE, DK, EE, ES, FI, FR, GB, GR, HR, HU, IE, IS, IT, LT, LU, LV, MC, MK, MT, NL, NO, PL, PT, RO, RS, SE, SI, SK, SM, TR)
African Intellectual Property Organization (BF, BJ, CF, CG, CI, CM, GA, GN, GQ, GW, KM, ML, MR, NE, SN, TD, TG)
Publication Language: Chinese (ZH)
Filing Language: Chinese (ZH)