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High-throughput screening method for fast-growing type acid-producing strains of ketogulonigenium vulgare

A technology of ketogenic cologne and screening method, which is applied in the direction of mutant preparation, etc. It can solve the problems of positive mutant strains without purpose, increase the number of culture devices, and large screening volume, etc., and achieve high-throughput screening and simple breeding , the effect of rapid identification

Inactive Publication Date: 2016-05-04
石药集团维生药业(石家庄)有限公司
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AI Technical Summary

Problems solved by technology

Such method has the following disadvantages: the one is through mutagenesis treatment, even if the bacterial strains whose acid production performance is changed have almost no change in phenotypic characteristics such as colony shape and thallus shape, it is difficult to identify, so there is no way to select positive mutant strains. Purpose, and in order to avoid missed selection, basically all the single colonies that survived the mutagenesis are selected, and the screening amount is huge; second, the culture container used for the growth and metabolism detection of the strain is a test tube or a triangle with a relatively large liquid volume and volume. bottle, the amount of the strains to be tested is also relatively large, and the single colony needs to be enriched and cultivated first, which not only increases the cultivation time by about 3 days, but also increases the cultivation equipment

Method used

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  • High-throughput screening method for fast-growing type acid-producing strains of ketogulonigenium vulgare
  • High-throughput screening method for fast-growing type acid-producing strains of ketogulonigenium vulgare
  • High-throughput screening method for fast-growing type acid-producing strains of ketogulonigenium vulgare

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Experimental program
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Embodiment 1

[0026] In Example 1, the indicator added to the solid medium for identification is a mixture of bromothymol blue, bromocresol violet, and methyl red added with an appropriate amount of bromocresol violet and methyl red.

Embodiment 2

[0027] In Example 2, the indicators added to the solid medium for identification and the seed liquid medium for identification are both bromothymol blue.

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Abstract

The invention provides a high-throughput screening method for fast-growing type acid-producing strains of ketogulonigenium vulgare, and belongs to the technical field of microbial strain breeding. According to the high-throughput screening method, the ketogulonigenium vulgare subjected to mutagenic treatment is cultured on a solid medium panel, a certain dose of an indicator is added to the solid medium, and simple and fast identification for the fast-growing type acid-producing strains of the ketogulonigenium vulgare is realized; according to design, the selected fast-growing type acid-producing strains of the ketogulonigenium vulgare are inoculated to a strain liquid medium for culturing, a certain dose of the indicator is added to the strain liquid medium, simple and fast determination for the selected fast-growing type acid-producing strains of the ketogulonigenium vulgare is realized, then the indicator identification is combined with a centrifuge tube group culture technology, and the batch culturing effect is realized while the requirement for independent sterile operation in single colony inoculation culture and detection processes of the ketogulonigenium vulgare is met, so that the purpose of rapid, convenient and accurate high-throughput screening is achieved.

Description

technical field [0001] The invention relates to a microbial strain breeding method, in particular to a high-throughput screening method for fast-growing acid-producing bacterial strains of common ketogenic cologne. Background technique [0002] Vitamin C is an essential vitamin for the human body. It has a wide range of physiological functions and plays an important role in the medicine and food industries. At present, the "two-step fermentation method" is widely used in China. The first step: single-bacteria fermentation to convert D-sorbitol into L-sorbose; Commonly known as small bacteria) and Bacillus megaterium (commonly known as large bacteria) to convert L-sorbose into 2-keto-L-gulonic acid; 2-keto-L-gulonic acid is then extracted and transformed into vitamin c. [0003] In the "two-step fermentation" process of producing vitamin C, mixed bacteria are fermented, and L-sorbose is converted into 2-keto-L-gulonic acid through the joint action of common ketogenic cologn...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/01
CPCC12N15/01
Inventor 焦迎晖胡静王清格马乐辉焦暖刘丽华安卫红李迎辉
Owner 石药集团维生药业(石家庄)有限公司
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