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1. WO2018149282 - METHOD FOR HETEROLOGOUS EXPRESSION OF EPOTHILONE

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Publication Number WO/2018/149282
Publication Date 23.08.2018
International Application No. PCT/CN2018/074229
International Filing Date 26.01.2018
IPC
C12P 17/18 2006.1
CCHEMISTRY; METALLURGY
12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
PFERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
17Preparation of heterocyclic carbon compounds with only O, N, S, Se, or Te as ring hetero atoms
18containing at least two hetero rings condensed among themselves or condensed with a common carbocyclic ring system, e.g. rifamycin
C12N 1/21 2006.1
CCHEMISTRY; METALLURGY
12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
NMICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
1Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
20Bacteria; Culture media therefor
21modified by introduction of foreign genetic material
CPC
C12N 1/145
C12N 1/205
C12N 15/52
CCHEMISTRY; METALLURGY
12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
NMICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
15Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
09Recombinant DNA-technology
11DNA or RNA fragments; Modified forms thereof
52Genes encoding for enzymes or proenzymes
C12P 17/18
CCHEMISTRY; METALLURGY
12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
PFERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
17Preparation of heterocyclic carbon compounds with only O, N, S, Se or Te as ring hetero atoms
18containing at least two hetero rings condensed among themselves or condensed with a common carbocyclic ring system, e.g. rifamycin
C12P 17/181
CCHEMISTRY; METALLURGY
12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
PFERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
17Preparation of heterocyclic carbon compounds with only O, N, S, Se or Te as ring hetero atoms
18containing at least two hetero rings condensed among themselves or condensed with a common carbocyclic ring system, e.g. rifamycin
181Heterocyclic compounds containing oxygen atoms as the only ring heteroatoms in the condensed system, e.g. Salinomycin, Septamycin
C12R 2001/01
Applicants
  • 复旦大学 FUDAN UNIVERSITY [CN]/[CN]
Inventors
  • 丁晓明 DING, Xiaoming
  • 禹育聪 YU, Yucong
  • 唐标 TANG, Biao
  • 张霖 ZHANG, Lin
  • 王慧敏 WANG, Huimin
  • 卞小莹 BIAN, Xiaoying
  • 李越中 LI, Yuezhong
  • 张友明 ZHANG, Youming
  • 赵国屏 ZHAO, Guoping
Priority Data
201710090318.720.02.2017CN
Publication Language Chinese (zh)
Filing Language Chinese (ZH)
Designated States
Title
(EN) METHOD FOR HETEROLOGOUS EXPRESSION OF EPOTHILONE
(FR) PROCÉDÉ D'EXPRESSION HÉTÉROLOGUE D'UN ÉPOTHILONE
(ZH) 一种异源表达埃博霉素的方法
Abstract
(EN) Disclosed is a method for heterologous expression of an epothilone, comprising introducing an epothilone gene cluster into host bacteria, while supplementing an epothilone precursor synthesis pathway combining with the introduction of the related tRNA gene and insertion of a promoter. The method can substantially increase the expression quantity of the epothilone, and the yield can be improved by about four orders of magnitude and can reach 8.5 mg/L. In addition, also provided are a genetic engineering strain for heterologous expression of the epothilone and a method for producing the epothilone.
(FR) L'invention concerne un procédé pour l'expression hétérologue d'un épothilone, comprenant l'introduction d'un groupe de gènes d'épothilone dans des bactéries hôtes, tout en complétant une voie de synthèse de précurseur d'épothilone avec l'introduction du gène d'ARNt associé et l'insertion d'un promoteur. Le procédé peut sensiblement augmenter la quantité d'expression de l'épothilone, et le rendement peut être amélioré par environ quatre ordres de grandeur et peut atteindre 8,5 mg/L. En outre, l'invention concerne également une souche d'ingénierie génétique pour l'expression hétérologue de l'épothilone et un procédé pour produire l'épothilone.
(ZH) 本发明公开了一种异源表达埃博霉素的方法，在宿主菌中引入埃博霉素基因簇，同时补充埃博霉素前体合成途径，结合相关 tRNA 基因的引入及启动子的插入，能够大幅提高埃博霉素的表达量，产量提高约 4 个数量级，可达到 8.5mg/L 。此外，本发明还提供了一种异源表达埃博霉素的基因工程菌株及生产埃博霉素的方法。
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