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
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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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