Method for synthesizing glyoxylic acid by utilizing corynebacterium glutamicum based on CRISPRi regulation

A technology of glutamic acid stick and glyoxylic acid, applied in the field of bioengineering, can solve few problems and achieve the effect of reducing the generation of by-products

Active Publication Date: 2021-07-20
HEBEI UNIVERSITY OF SCIENCE AND TECHNOLOGY
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Problems solved by technology

[0006]It is very interesting for researchers to use CRISPRi technology to regulate the synthesis of glyoxylic acid, so as to help realize the industrial production of glyoxylic acid by biological methods Significance, but there are few reports about the production of glyoxylic acid by fermentation of genetically engineered bacteria in the prior art

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  • Method for synthesizing glyoxylic acid by utilizing corynebacterium glutamicum based on CRISPRi regulation
  • Method for synthesizing glyoxylic acid by utilizing corynebacterium glutamicum based on CRISPRi regulation
  • Method for synthesizing glyoxylic acid by utilizing corynebacterium glutamicum based on CRISPRi regulation

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Embodiment Construction

[0050] It should be noted that, in the case of no conflict, the embodiments of the present invention and the features in the embodiments can be combined with each other.

[0051] The present invention will be described in detail below with reference to the accompanying drawings and examples.

[0052] This example relates to a method for synthesizing glyoxylic acid using Corynebacterium glutamicum based on CRISPRi regulation.

[0053] Corynebacterium glutamicum ATCC 13032 (C. glutamicum ATCC 13032 for short), as a food-safe microorganism, has the advantages of easy cultivation, no spore production, high yield, and biosafety. With the completion of the whole genome sequencing of Corynebacterium glutamicum and the accumulation of various omics data, the directional transformation and construction of engineering strains have been realized. During the glycolytic pathway and TCA cycle of C. glutamicum, pyruvate, lactate There is a complex network system among citric acid, isocitric...

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Abstract

The invention provides a method for synthesizing glyoxylic acid by utilizing corynebacterium glutamicum based on CRISPRi (clustered regularly interspaced short palindromic repeats i) regulation. According to the method, an acetaldehyde dehydrogenase coding gene of the corynebacterium glutamicum is replaced through a dCas9 expression cassette gene; a lactic dehydrogenase encoding gene of corynebacterium glutamicum is replaced by sgRNA transcription units of an isocitrate dehydrogenase encoding gene and a malic acid synthetase encoding gene, an isocitrate lyase encoding gene of the corynebacterium glutamicum is over-expressed to obtain a corynebacterium glutamicum engineering strain, and the engineering strain is cultured to ferment and synthesize glyoxylic acid. Corynebacterium glutamicum is used as a starting strain, a lactic dehydrogenase coding gene and an acetaldehyde dehydrogenase coding gene which are key enzymes of branch metabolism of the corynebacterium glutamicum are knocked out, a CRISPRi regulation system is established, expression levels of an isocitrate dehydrogenase coding gene and a malic acid synthase coding gene are reduced, and meanwhile, an isocitrate lyase coding gene is overexpressed, so that a method for biosynthesizing glyoxylic acid by using corynebacterium glutamicum is established.

Description

technical field [0001] The invention relates to the technical field of bioengineering, in particular to a method for synthesizing glyoxylic acid using Corynebacterium glutamicum based on CRISPRi regulation for glyoxylic acid biosynthesis. Background technique [0002] Glyoxylic acid (Glyoxylic acid) is the substance with the smallest molecular weight in the alkydric acid family. It is an important organic acid in the TCA cycle (tricarboxylic acid cycle) and glyoxylic acid cycle of organisms, which affects the metabolic balance of organisms. Its derivatives There are dozens of kinds, among which the typical ones are vanillin, ethyl vanillin, allantoin, p-hydroxyphenylalanine, p-hydroxyphenylacetamide and so on. Glyoxylic acid derivatives are widely used in food additives, cakes, candy, baked food, tobacco and alcohol, spices, cosmetics and pharmaceutical industries. With the increase in demand for glyoxylic acid, the chemical production of glyoxylic acid brings Pollution is ...

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/10C12N15/77C12N15/66C12P7/40C12R1/15
CPCC12N15/102C12N15/77C12N15/66C12P7/40
Inventor 冯惠勇李天明王丽丽
Owner HEBEI UNIVERSITY OF SCIENCE AND TECHNOLOGY
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