Method for screening high-yield bacterial strain of vitamin B2

A technology of high-yield strains and screening methods, applied in the field of bioengineering, can solve the problems of unstable strains, large randomness, and no repeatability, and achieve the effects of reducing fermentation costs, improving synthesis capacity, and overcoming low yields

Inactive Publication Date: 2017-12-15
SHANGHAI ACEBRIGHT PHARMA CO LTD +2
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Problems solved by technology

This kind of conventional mutagenesis is quite random, and the experimental results are almost non-repeatable, and the mutagenized strain is unstable, and it is easy to restore the mutation after passage, resulting in the instability of the strain and adversely affecting the subsequent screening of the strain. , which is not conducive to increasing the production of vitamin B2

Method used

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Examples

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Embodiment

[0025] A screening method for vitamin B2 high-yield strains, comprising the steps of:

[0026] a) Pretreatment of the original bacterial strain: 80 μl of the glycerol bacteria of the original bacterial strain of Bacillus subtilis (Bacillus subtilis) were evenly spread on the plate medium and cultivated at 37°C for 24 hours. The components of the plate medium were: yeast powder 5g / L, sodium chloride 10g / L, tryptone 10g / L, agar powder 18g / L, pH7.0; Bacteria, after mixing evenly, draw into a sterile centrifuge tube to obtain the bacterial liquid of the original bacterial strain of Bacillus subtilis, and set aside;

[0027] b) Take 2ml of the bacterial solution obtained in step a) and drop it on a sterile blank culture plate, and perform ultraviolet mutagenesis under sterile conditions for 30s to obtain an ultraviolet mutagenized bacterial strain;

[0028] c) Take 0.2ml of the bacterial liquid of the ultraviolet mutagenic strain obtained in step b), add it to 0.1mol / L ethyl meth...

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Abstract

The invention discloses a method for screening high-yield vitamin B2 strains. The vitamin B2 high-yield strains are obtained through the following steps: the original strains are pretreated, subjected to ultraviolet and chemical mutagenesis, and then domesticated by metabolic substrate analogues, and finally re-treated. It is obtained by screening nutrient medium and fermentation medium. The present invention comprehensively utilizes physical and chemical compound mutagenesis and domestication of metabolic substrate analogues, thereby screening out a stable and high-yielding vitamin B2 strain, which overcomes the defect of low vitamin B2 production in the prior art compared with the original strains , under the condition that the fermentation cycle remains unchanged, its vitamin B2 synthesis ability is increased by about 6g / L, and the fermentation cost is greatly reduced.

Description

technical field [0001] The invention relates to a screening method for bacterial strains, in particular to a screening method for high-yield vitamin B2 bacterial strains, and belongs to the technical field of bioengineering. Background technique [0002] Vitamin B2 (chemical formula: C17H20N4O6, formula weight 376.37), also known as riboflavin, is a water-soluble B vitamin. Vitamin B2 is one of the essential vitamins for humans and animals. It is a component of flavin enzyme coenzymes and mainly exists in the form of flavin mononucleotide (FMN) and flavin adenine dinucleotide (FAD) in organisms. , participate in the electron transfer and redox reaction of the respiratory chain of the body tissue, play an important role in respiration and biological oxidation, and are indispensable vitamins in life activities. [0003] At present, the production methods of vitamin B2 used in industry mainly include chemical synthesis, chemical semi-synthesis and microbial fermentation. Comp...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N13/00C12N15/01C12N1/36C12R1/125
CPCC12N13/00C12N1/36C12N15/01
Inventor 张国银孙向宇
Owner SHANGHAI ACEBRIGHT PHARMA CO LTD
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