Technique for producing gene rHu(Cu/Zn-SOD) with one-step method
A technology of superoxide dismutase and genetic recombination, applied in the direction of enzymes, biochemical equipment and methods, enzymes, etc., can solve problems such as difficult industrial mass production, limited raw material sources, and complicated extraction process, and achieve low unit cost and no The effect of residual virus and process simplification
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[0040] This process is implemented and applied in a 100L fermentation tank.
[0041] 1. Preparation of seed solution
[0042] 1) Take a frozen engineered bacteria (PBV / STII / SOD), use an inoculation loop to take a small amount of strains, draw a line on an LB plate, and culture at 30°C for 24 hours.
[0043] 2) Pick a single colony and inoculate it into an LB medium test tube (LB culture medium: 56g of peptone, 28g of yeast powder, 56g of NaCe, 5.6L of fixed solution) and sterilize it, then add a small amount of ampicillin, and put it in a constant temperature shaker for cultivation at 30°C. 170 rpm for 10-24 hours, OD 600 When it is 0.4-0.6, it is connected to the fermenter.
[0044] 2. Fermentation tank culture
[0045] 1) Fermentation broth adopts improved M 9 Medium (M 9 Preparation of medium: 800g peptone, 500g yeast powder, NaHPO 4 320g, KH 2 PO 4 110g, NaCe100g, NH 4 Ce70g, MgSo 4 13.4g, CaLe0.5g, CuSo 4 10g, ZnSo 4 0.5g, glucose 600g. )
[0046] 2) After t...
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