人胰高血糖素样肽-1突变体融合蛋白在大肠杆菌中的自诱导表达优化

考察了在大肠杆菌中自诱导表达人胰高血糖素样肽-1突变体融合蛋白的可行性,并对自诱导培养条件及培养基成分进行优化,以提高蛋白产量。实验结果表明,最优培养基成分为蛋白胨19.17g/L,酵母膏9.59g/L,Na2HPO45.72g/L,KH2PO45.48g/L,(NH4)2SO42.66g/L,NaCl3.33g/L,甘油2%(V/V),葡萄糖0.68g/L,乳糖6.33g/L,MgSO40.24g/L。在温度33°C、接种量1%、pH7、装瓶量20mL/100mL培养条件下,用该最优培养基自诱导表达人胰高血糖素样肽-1突变体融合蛋白的产量可达348.6mg/L。

Auto-inducing Expression Optimization of Mutant Human Glucagon-like Peptide-1 Fusion Protein in Escherichia coli BL21 (DE3)

The auto-inducing expression of mutant human glucagon-like peptide-1 fusion protein in Escherichia coli BL21 (DE3) was studied. In order to increase the protein yield, we optimized both the auto-induction fermentation conditions and medium. The results showed that the optimal auto-induction medium is: tryptone 19.17 g/L, yeast extract 9.59 g/L, Na2HPO4 5.72 g/L, KH2PO4 5.48 g/L, (NH4)2SO4 2.66 g/L, NaCl 3.33 g/L, glycerol 2% (v/v), glucose 0.68 g/L, lactose 6.33 g/L, MgSO4 0.24 g/L. The genetically engineered strain was cultivated at 33°C, pH7, in a 100 mL flask containing 20 mL such medium, with the inoculum size of 1%, for 12 h. Then the yield of mutant human glucagon-like peptide-1 fusion protein reached 348.6 mg/L.