重组人基质金属蛋白酶2(MMP-2)血红素样蛋白表达优化

Optimization of expression conditions of recombinant human-source matrix metalloproteinase 2 hemopexin-like C-terminal domain

为确定重组大肠杆菌(BL21(DE3)/PET42a-PEX)最佳表达条件。在单因素实验的基础之上,以目的蛋白PEX表达量为响应值,应用Box-Behnken中心组合设计建立数学模型,进行响应面分析。响应面分析确定的最优因素水平组合为:LB培养基pH为7.50,诱导时菌液OD600为0.60,IPTG诱导浓度为0.60mmol/L,诱导培养时间为5.0h。对此工艺条件进行验证得到目的蛋白PEX表达量为20.98mg/L。结果显示,优化了基因重组大肠杆菌表达人PEX的条件,为实现基因重组原核工程菌发酵法制备重组PEX奠定了基础。

To optimize fermentation conditions of Genetically Engineered Escherichia coli（BL21（DE3）/PET42a-PEX）,Box-Behnken design was used to establish mathematical model based on single-factor tests.The response surface methodology（RSM） analysis result showed that the optimal scale of factors was：Luria-Bertani liquid culture medium pH7.50,bacterial liquid optical density（OD600） 0.60,induce concentration of IPTG 0.60mmol/L,induce time 5.0h.Under these optimized conditions,the yield of PEX was 20.98mg/L.Optimization of expression conditions of genetically engineered Escherichia coli expressing recombinant human-source matrix metalloproteinase 2 hemopexin-like C-terminal domain was studied,and laid the foundation of generation of human recombinant PEX by fermentation.