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本研究对影响FGF21(L~(59)R)突变体基因工程菌发酵条件的因素进行了优化。采用正交试验确定FGF21(L~(59)R)突变体基因工程菌的最佳LB培养基配方,利用SDS-PAGE电泳检测不同发酵条件对FGF21(L~(59)R)突变体蛋白表达情况。LB培养基最佳配比(g/L):蛋白胨11,酵母粉6,氯化钠10,葡萄糖1;在此基础上优化基因工程菌发酵条件,确定LB培养基p H 6.5~7.2,装液量(溶解氧)20%,菌体密度(A_(600))1.0,IPTG浓度0.6 mmol/L,37℃条件下诱导5 h,突变体蛋白的表达量由优化前的12%提高至35%。结果表明,培养基配方、p H、装液量(溶解氧)、菌体密度(A_(600))、IPTG浓度、温度、诱导时间均对FGF21(L~(59)R)突变体基因工程菌表达量有影响。
In this study, the factors influencing the fermentation conditions of genetically engineered strains of FGF21 (L ~ (59) R) mutant were optimized. The optimal LB medium of FGF21 (L ~ (59) R) mutant was identified by orthogonal test. The protein expression of FGF21 (L ~ (59) R) mutant was detected by SDS-PAGE. Happening. LB medium optimized ratio (g / L): peptone 11, yeast powder 6, sodium chloride 10, glucose 1; on this basis, to optimize the fermentation conditions of genetically engineered bacteria to determine LB medium p H 6.5 ~ 7.2, The optimal conditions were as follows: volume of solution (dissolved oxygen) 20%, cell density (A_ (600)) 1.0, IPTG concentration 0.6 mmol / L and induction at 37 ℃ for 5 h. %. The results showed that the effects of medium formulation, p H, liquid loading (dissolved oxygen), cell density (A 600), IPTG concentration, temperature and induction time on the genetic engineering of FGF21 (L ~ (59) The amount of bacteria affected.