重组脂肪酶自诱导发酵培养基优化

Optimization of auto-induction fermentation medium for recombined lipase

关键词:  脂肪酶  自动诱导  响应面  培养基优化

Key words:  lipase  auto-induction  response surface method  medium optimization

摘要:

[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,KH₂PO₄ 0.68,NH₄Cl 0.267 5,Na₂SO₄ 0.071,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=6.35 U/mg,比初始发酵培养基提高近3倍。

Abstract:

[Objective] Optimizing the auto-induction of the fermentation medium to increase lipase enzyme production.[Methods] In this research, through the construction of a transposon-integrated lipase recombinant bacterial strain, using automatic induction fermentation expression 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 production.[Results] The optimized fermentation medium (W/V) contained glycerol 2.596, glucose 0.035, lactose 1.289, tryptone 1.0, yeast extract 0.5, Na₂HPO₄·12H₂O 1.79, KH₂PO₄ 0.68, NH₄Cl 0.267 5, Na₂SO₄0.071, MgSO₄·7H₂O 0.05.[Conclusion] The results of the fermentation test confirmed that the lipase enzyme specific activity in the optimized fermentation medium was 6.35 U/mg, which was nearly three times higher than that in the initial fermentation medium.