Method for producing 5-glycyl ethylformic acid by using engineering bacterium
A technology of aminolevulinic acid and engineering bacteria, which is applied in the directions of microorganism-based methods, biochemical equipment and methods, microorganisms, etc., can solve the problems of complex culture conditions, many by-products, and complicated steps, and achieves broad industrialization prospects. The effect of high controllability and simple process
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[0018] The method for producing 5-aminolevulinic acid with engineering bacteria comprises the following steps:
[0019] 1. Use an inoculation needle to dip a small amount of bacterial liquid from the glycerol tube of CGMCC No.1939 engineering bacteria Rosetta (DE3)-pET28a-R.S.hemA, and add it to the chloramphenicol containing 30 μg / ml kanamycin and 34 μg / ml chloramphenicol. Streak on the LB medium plate and cultivate overnight in a 37°C oven;
[0020] 2. Pick a single clone and inoculate it in a 250ml shake flask containing 50ml LB medium at a rotation speed of 200rmp, and culture overnight at 37°C to obtain first-grade seeds;
[0021] 3. Take 1ml of primary seeds and inoculate them in a 500ml shake flask containing 100ml of fermentation medium, culture at 37°C and 200rpm for 3h to obtain secondary seeds. Wherein the initial fermentation medium is composed of 1% peptone, 0.5% yeast powder, 0.3% succinic acid, 0.2% glycine and 0.2% glucose in mass volume ratio, and the pH valu...
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