Construction method and application of genetically engineered bacterium for high-yield of cephalosporin C

A technology of genetically engineered bacteria and cephalosporins, applied in the direction of genetic engineering, microbial-based methods, applications, etc., can solve the problems of oil residues, limited lipase, etc., to improve utilization efficiency, promote synthesis rate and yield, and be good for industry The effect of applying the foreground

Active Publication Date: 2021-07-13
河南省健康元生物医药研究院有限公司
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Problems solved by technology

In previous studies, it was found that the lipase synthesized by the wild-type C. chrysogenum was limited, and there was a large amount of oil remaining in the fermentation broth after the fermentation

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  • Construction method and application of genetically engineered bacterium for high-yield of cephalosporin C
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  • Construction method and application of genetically engineered bacterium for high-yield of cephalosporin C

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Embodiment Construction

[0024]The present invention will be further described below in conjunction with the accompanying drawings and specific embodiments. The schematic embodiments and descriptions of the present invention are used to explain the present invention, but are not intended to limit the present invention.

[0025] Bacterial strains and plasmids used in this embodiment: Escherichia coli E. coli DH5α (purchased from Takara, Japan); the strain of the overexpressed lipase gene is Cephalosporium chrysoprase ( Acremonium chrysogenum ) ATCC11550 and CGMCC3.3795, which are stored in American type culture collection (ATCC) and China General Microbiological Culture Collection Center (CGMCC) respectively. Plasmid pBARGPE1-Hygro is a general-purpose filamentous fungal expression plasmid, which was purchased from Miaoling Plasmid Platform (Product No.: P0381). For the construction and operation method of the plasmid pBARGPE1-ACLipaseOE, please refer to the literature (Song Jia. Preliminary explora...

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Abstract

The invention discloses a construction method and application of a genetically engineered bacterium for high-yield of cephalosporin C, and relates to the technical field of preparation of cephalosporin C. According to the construction method, an endogenous lipase gene of acremonium chrysogenum is over-expressed, so that the lipase activity in a CPC fermentation process is improved, triglyceride in soybean oil is promoted to be hydrolyzed into molecules such as fatty acid, so that the fermentation yield of CPC is increased. The lipase gene AclipA of acremonium chrysogenum disclosed by the invention can effectively regulate the synthesis rate and yield of cephalosporin C of acremonium chrysogenum. By utilizing the lipase gene AclipA of acremonium chrysogenum and the protein coded thereby, the utilization efficiency of acremonium chrysogenum bacterium on fermentation raw material soybean oil can be effectively improved, the synthesis rate and the yield of cephalosporin C are promoted, and the method has a very good industrial application prospect.

Description

technical field [0001] The invention relates to the technical field of genetically engineered bacteria, in particular to a method for constructing genetically engineered bacteria with high cephalosporin C production and its application. Background technique [0002] Cephalosporin C (CPC) is the second β-lactam antibiotic found in nature, and it is also the main raw material for the production of 7-aminocephalosporanic acid (7-ACA), an important intermediate of cephalosporin antibiotics. CPC can inhibit bacterial cell wall synthesis, but its antibacterial activity is low. At the same time, it has gained widespread attention and research because it is safe and non-toxic to the human body. At present, the industry mainly utilizes Cephalosporium acremonium ( Acremonium chrysogenum ) to produce CPC by fermentation. After years of research, the biosynthesis pathway of CPC in Cephalosporium chrysogenum has been clearly studied, and the genetic operating system of Cephalosporium ...

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Application Information

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IPC IPC(8): C12N1/15C12N15/57C12N9/20C12N15/80C12P35/06C12R1/645
CPCC12N9/20C12N15/80C12P35/06C12Y301/01003
Inventor梁恒宇韩超熊琼超李雪亮王世超周鹏
Owner河南省健康元生物医药研究院有限公司