A kind of isopropylmalate synthase and its application
A malate synthase, isopropyl technology, applied in the field of metabolic engineering, can solve the problems of unstable fermentation performance, slow growth, unstable genetic traits, etc.
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[0020] Example 1: The gene leuA encoding 2-isopropylmalate synthase that relieves L-leucine feedback inhibition M the acquisition
[0021] (1) Screening of mutants resistant to L-leucine structural analogues
[0022] ① Preparation of Corynebacterium glutamicum (Corynebacterium glutamicum) ATCC13032 bacterial suspension
[0023] Corynebacterium glutamicum (Corynebacterium glutamicum) ATCC13032 was inoculated into LB liquid medium, cultured at 32°C, 200rpm for 12h, collected by centrifugation, washed with sterile saline for 3 times and then resuspended to make the OD 600 =0.6-0.8, take 10 μL of the bacterial suspension and spread it on the slide.
[0024] ② Plasma mutagenesis at atmospheric pressure and room temperature
[0025] The mutagenesis parameters were as follows: the slide was placed at 2mm from the airflow port, the power was 120W, the airflow was 10SLM, and the action time was 20s.
[0026] ③ Screening of mutant strains resistant to L-leucine structural analog leu...
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