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Construction method of monascus genetically engineered microorganism having high-yield product Monacolin K and application of monascus genetically engineered microorganism

A technology of genetically engineered bacteria and construction methods, which is applied in the field of microbial genetic engineering, can solve the problem of low yield of MonacolinK, and achieve the effect of convenient method and strong ability

Inactive Publication Date: 2017-07-28
BEIJING TECHNOLOGY AND BUSINESS UNIVERSITY
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Problems solved by technology

[0006] Aiming at the deficiencies in the prior art, the present invention provides a method for constructing MonacolinK genetically engineered bacteria with high yield of MonacolinK and its application. Using the method of the present invention, the metabolic pathway of MonacolinK in MonacolinK can be enhanced, and then fundamentally solve the problem of MonacolinK. The problem of low yield of Monacolin K among super metabolites

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  • Construction method of monascus genetically engineered microorganism having high-yield product Monacolin K and application of monascus genetically engineered microorganism
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  • Construction method of monascus genetically engineered microorganism having high-yield product Monacolin K and application of monascus genetically engineered microorganism

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Embodiment 1

[0051] Embodiment 1, the construction of Monascus genetically engineered bacteria with high yield of MonacolinK

[0052] 1. Overexpression plasmid construction technical route

[0053] The MonacolinK high-yield Monascus genetically engineered bacteria of the present invention is obtained by using the pBC-Hygro plasmid as an overexpression plasmid and overexpressing mokC or mokD or mokE or mokI genes. The route is: extract Monascus RNA, reverse transcribe it into cDNA, and use the cDNA as a template to amplify the target fragment. After the target fragments are double-digested and connected, the recombinant plasmid is constructed. Taking pBC-hygro-mokC as an example, the technical route is as follows: figure 1 shown.

[0054] 2. Extraction of Monascus RNA

[0055] Purify the mycelia of Monascus violaceum M1 after fermentation and cultivation to the 8th day, and then extract the transcriptome RNA. The extraction method is a conventional method in the art. Take 7 μL RNA sampl...

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Abstract

The invention provides a construction method of monascus genetically engineered microorganism having high-yield product Monacolin K and application of the monascus genetically engineered microorganism. Monacolin K encoding gene, namely mok gene, is subjected to overexpression in monascus so that the monascus genetically engineered microorganism having high-yield product Monacolin K is obtained. The mok gene refers to one of mok C gene, mok D gene, mok E gene and mok I gene. Metabolic pathways of Monacolin K of monascus can be increased, and thus, the problem about low yield of secondary metabolite, namely Monacolin K of monascus can be solved fundamentally. Besides, the genetically engineered microorganism is capable of reducing content of citrinin and achieving the purposes of improving quality of monascus products and guaranteeing safety of the monascus products.

Description

technical field [0001] The invention belongs to the technical field of microbial genetic engineering, and relates to a construction method and an application of Monacolin K high-yield Monascus genetically engineered bacteria. Background technique [0002] Monascus purpureus Went. (Monascus purpureus Went.), also known as red yeast rice, red rice, red rice, exists in trees, soil and accumulations. It is an important microbial resource in my country, which can produce a wide range of natural products with biological activity and has high commercial value. Monascus can grow in a wide temperature range, the general temperature is 15-42°C, the optimum temperature is 25-30°C, the optimum pH value for growth is 3.5-5, and it can tolerate ethanol with a concentration of up to 10%. It grows well on malt juice agar medium, and the colony is white at first, and becomes light pink, purple or gray-black when mature, and often turns red. [0003] The application of red yeast rice has a ...

Claims

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Application Information

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IPC IPC(8): C12N1/15C12N15/31C12N15/80C12P17/06C12R1/645
CPCC07K14/37C12N15/80C12P17/06
Inventor 张婵王成涛孙宝国梁健柴诗缘
Owner BEIJING TECHNOLOGY AND BUSINESS UNIVERSITY
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