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Escherichia coli and application thereof in fermentation production of L-tryptophan

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

Pending Publication Date: 2022-05-13
江苏澳创生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, strains have high requirements on raw materials, culture conditions and control technology, 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 tryptophan, but L-tryptophan 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 and application thereof in fermentation production of L-tryptophan
  • Escherichia coli and application thereof in fermentation production of L-tryptophan

Examples

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Embodiment 1

[0021] Example 1: A strain of Escherichia coli and its application in fermentative production of L-tryptophan

[0022] A strain of Escherichia coli ACTrp104, which is classified as Escherichia coli (Escherichia coli), was preserved in the General Microbiology Center (CGMCC) of the China Committee for the Collection of Microorganisms, and the preservation time was December 03, 2018. The species deposit number is CGMCC No.16861.

[0023] The Escherichia coli ACTrp104 and its application in fermentative production of L-tryptophan comprise the following steps:

[0024] (1) Strain culture, sterilize the seed medium, cool it to about 35°C, adjust the pH to about 7, inoculate the Escherichia coli ACTrp104 strain into the seed medium for cultivation, and cultivate to logarithmic growth Expect. Bacteria culture conditions: temperature 35°C, pH value around 7, dissolved oxygen ≥ 30%, tank pressure 0.03MPa.

[0025] (2) Fermentation culture, the fermentation medium is sterilized, cool...

Embodiment 2

[0033] Example 2: A strain of Escherichia coli and its application in fermentative production of L-tryptophan

[0034] A strain of Escherichia coli ACTrp104, which is classified as Escherichia coli (Escherichia coli), was preserved in the General Microbiology Center (CGMCC) of the China Committee for the Collection of Microorganisms, and the preservation time was December 3, 2018. The species deposit number is CGMCC No.16861.

[0035] The Escherichia coli ACTrp104 and its application in fermentative production of L-tryptophan comprise the following steps:

[0036] (1) Strain culture, sterilize the seed medium, cool it to about 30°C, adjust the pH to about 6.5, inoculate the Escherichia coli ACTrp104 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.

[0037](2) Fermentation culture, the fermentation medium is sterilized, c...

Embodiment 3

[0045] Example 3: A strain of Escherichia coli and its application in fermentative production of L-tryptophan

[0046] Escherichia coli mutagenesis screening method

[0047] Using Escherichia coli AC-1042 as the starting bacterium, through atmospheric and room temperature plasma (ARTP) mutagenesis treatment, and then through the directed selection of structural analog resistance, tryptophan with substrate resistance and genetic markers was obtained High-yielding strain. ARTP mutagenesis treatment method: Pick a ring of bacteria from the freshly activated slant and put it in the seed medium, culture it on a shaker at 180r / min at 36.5°C for 4.5h, take 1mL of the seed solution in a 1.5mL EP tube, centrifuge at 4000rpm, remove the above To the supernatant, add 1mL of normal saline, mix well, repeat three times, dilute the bacterial suspension to make the bacterial suspension OD 600 0.6-1.0; take 10 μL of diluted bacterial solution and spread it evenly on a sterile stainless stee...

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Abstract

The invention relates to Escherichia coli and an application thereof in fermentation production of L-tryptophan, the Escherichia coli is Escherichia coli ACTrp104, the Escherichia coli ACTrp104 is preserved in the China General Microbiological Culture Collection Center (CGMCC), the preservation date is December 3, 2018, and the preservation number of the strain is CGMCC No.16861. The invention further discloses an application of the Escherichia coli in fermentation production of L-tryptophan. The Escherichia coli AC-1042 is screened by combining an ARTP mutation breeding technology and a resistance directional screening technology, and the strain has unique physiological and biochemical characteristics and can be researched and applied in the fields of scientific research, industry and the like.

Description

technical field [0001] The invention relates to the field of biological technology, and specifically relates to a strain of Escherichia coli and its application in fermentative production of L-tryptophan. Background technique [0002] L-tryptophan is one of the essential amino acids in humans and animals, and is an important part of protein. Humans and animals cannot synthesize it by themselves and must be ingested from food. L-tryptophan is mainly used in medicine, food, cosmetics, feed and other industries. The latest research shows that tryptophan and its metabolites play different biological effects in the regulation of many human diseases, such as nervous system diseases, Tumor, nephropathy, diabetes, inflammation, immune disease, etc. In the field of food and feed additives, tryptophan can be used to strengthen food and make feed additives, and it plays an important role in supplementing the nutrition in food and feed and improving the utilization rate of protein. eff...

Claims

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Application Information

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IPC IPC(8): C12N1/20C12N13/00C12N15/01C12P13/22C12R1/19
CPCC12N1/20C12N13/00C12N15/01C12P13/227
Inventor 周旭波王健方培新王华萱
Owner 江苏澳创生物科技有限公司
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