Escherichia coli ACThr1032 and application thereof in fermentation production of L-threonine

A technology of Escherichia coli and threonine, applied in the direction of fermentation, microbial-based methods, bacteria, etc., can solve the problems of plasmid loss, genetic instability of strains, large raw materials and energy consumption, etc., to achieve acid production and conversion rate improvement, good industrial application prospects, and the effect of strain stability improvement

Active Publication Date: 2019-01-25
江苏澳创生物科技有限公司
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] At present, strains have high requirements on raw materials, culture conditions and control techniques, and the heredity of strains is also unstable in the production process, which makes the level of fermentation production unstable, acid production and conversion rate fluctuate greatly, resulting in raw materials and energy consumption. Larger, making the production cost higher
Through gene expression, the target gene is inserted into the plasmid to increase the plasmid with a larger gene copy number to increase the production of threonine, but L-threonine is a primary metabolite. During the cultivation process, the genetically modified microbial strains , there are many cases of plasmid loss and genetic instability, which are also a cause of fluctuations in production levels

Method used

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  • Escherichia coli ACThr1032 and application thereof in fermentation production of L-threonine
  • Escherichia coli ACThr1032 and application thereof in fermentation production of L-threonine

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Embodiment 1

[0022] The mutagenesis screening method of embodiment 1 escherichia coli

[0023] Using Escherichia coli E.coli THRD as the starting bacterium, after protoplast ultraviolet mutagenesis, diethyl sulfate (DES) chemical mutagenesis, nitrosoguanidine (NTG) chemical mutagenesis, and then separated and purified, a substance with substrate resistance was obtained. Sexually and genetically marked threonine-producing strains.

[0024] Protoplast UV mutagenesis method: Take 2-3 mL of the prepared protoplasts, add them to a plate with a diameter of 5 cm, place it under a 20w UV lamp, irradiate it vertically for 60-90 seconds, and then use a pipette gun to draw 0.2 mL and spread it on a petri dish medium, protected from light and incubated at 28-35°C for 36-72 hours.

[0025] Diethyl sulfate (DES) chemical mutagenesis method: After a seed culture with a threonine high-yielding slant strain, centrifuge for 10 minutes (3000-5000r / min), collect the bacteria, and wash the bacteria with steri...

Embodiment 2

[0036] A strain of Escherichia coli ACThr1032, which is classified as Escherichia coli, was deposited in the General Microbiology Center (CGMCC) of the China Committee for the Collection of Microorganisms, and the preservation date was July 23, 2018. The species deposit number is CGMCC No.16144.

[0037] The Escherichia coli ACThr1032 and its application in fermentative production of L-threonine comprise the following steps:

[0038] (1) Strain culture, sterilize the seed medium, cool it to about 30°C, adjust the pH to about 6.5, inoculate the Escherichia coli ACThr1032 strain into the seed medium for cultivation, and cultivate to logarithmic growth Expect. Bacterial culture conditions: temperature 30°C, pH value around 6.5, dissolved oxygen ≥ 30%, tank pressure 0.02MPa.

[0039] (2) Fermentation culture, the fermentation medium is sterilized, cooled to about 37°C, and the pH is adjusted to about 7.0, and the logarithmic growth phase bacterial strain obtained in step (1) is in...

Embodiment 3

[0047] A strain of Escherichia coli ACThr1032, which is classified as Escherichia coli, was deposited in the General Microbiology Center (CGMCC) of the China Committee for the Collection of Microorganisms, and the preservation date was July 23, 2018. The species deposit number is CGMCC No.16144.

[0048] The Escherichia coli ACThr1032 and its application in fermentative production of L-threonine comprise the following steps:

[0049] (1) Strain culture, sterilize the seed medium, cool it to about 40°C, adjust the pH to about 7.5, inoculate the Escherichia coli ACThr1032 strain into the seed medium for cultivation, and cultivate to logarithmic growth Expect. Bacterial culture conditions: temperature 40°C, pH value around 7.5, dissolved oxygen ≥ 30%, tank pressure 0.05MPa.

[0050] (2) Fermentation culture, the fermentation medium is sterilized, cooled to about 37°C, and the pH is adjusted to about 7.0, and the logarithmic growth phase bacterial strain obtained in step (1) is ...

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Abstract

The invention relates to a strain of Escherichia coli ACThr1032 and application thereof in fermentation production of L-threonine, and belongs to the field of biotechnology. The strain of Escherichiacoli ACThr1032 has a classification name of namedEscherichia coli, a deposit number of CGMCC No.16144, and a preservation date of July 23, 2018. The Escherichia coli ACThr1032 is cultured, fermented,purified and fermented to prepare L-threonine. The strain stability of Escherichia coli ACThr1032 in the invention is greatly improved, and the strain is used for fermentation production of L-threonine, acid production and conversion have been significantly improved, and breakthroughs have been made in technology, with significant progress. Moreover, the culture condition of the strain is extensive, which greatly reduces the energy consumption and has a good prospect of industrial application.

Description

technical field [0001] The invention relates to an Escherichia coli ACThr1032 and its application in fermenting and producing L-threonine, belonging to the field of biotechnology. Background technique [0002] L-threonine is an essential amino acid, which is mainly used in medicine, food, cosmetics, feed and other industries, especially in the rapid growth of feed additives. It is often added to the feed of immature piglets and poultry. It is the second limiting amino acid in pig feed and the third limiting amino acid in poultry feed. In recent years, the market demand for threonine has been increasing, and the market prospect is promising. [0003] At present, the production methods of threonine mainly include protein hydrolysis, chemical synthesis and microbial fermentation. Compared with protein hydrolysis and chemical synthesis, biological fermentation has been widely used in industrial production because of its environmental protection, high yield and high efficiency....

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20C12P13/08C12R1/19
CPCC12P13/08C12N1/205C12R2001/19
Inventor 周旭波王健方培新王华萱何亚章
Owner 江苏澳创生物科技有限公司
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