重组脂肪酶自诱导发酵培养基优化

摘要

[Objective]Optimizing the auto-induction of the fermentation medium to increase li-pase enzyme production.[Methods]In this research,through the construction of a transposon-integrated lipase recombinant bacterial strain,using automatic induction fermentation expres-sion method combined with CCD response surface experimental design method,the carbon source in the auto-induction medium was optimized,and a quadratic model was obtained,which was used to describe the effect of the carbon source in the fermentation medium on lipase pro-duction.[Results]The optimized fermentation medium(W/V)contained glycerol 2.596,glu-cose 0.035,lactose 1.289,tryptone 1.0,yeast extract 0.5,Na2HPO4·12H2O 1.79,KH2PO4 0.68,NH4Cl 0.2675,Na2SO40.071,MgSO4·7H2O 0.05.[Conclusion]The results of the fermentation test confirmed that the lipase enzyme specific activity in the optimized fermen-tation medium was 6.35 U/mg,which was near-ly three times higher than that in the initial fer-mentation medium.%[目的]优化自动诱导发酵培养基,提高脂肪酶产量.[方法]通过对构建的一株转座子整合型脂肪酶重组菌株,采用自动诱导发酵表达方法,并结合CCD响应面实验设计方法,对自动诱导培养基中的碳源进行优化,获得了 1 个二次模型用于描述发酵培养基中碳源对产脂肪酶的影响.[结果]优化后的最适自动诱导培养基(W/V)组成为 glycerol 2.596,glucose 0.035,lactose 1.289,tryptone 1.0,yeast extract 0.5,Na2HPO4·12H2O 1.79,KH2PO40.68,NH4Cl 0.267 5,Na2SO40.071,MgSO4·7H2O 0.05.[结论]发酵试验表明,最优碳源培养基的比活力为R1=6.3 5 U/mg,比初始发酵培养基提高近 3 倍.