Box-Behnken实验设计及响应面分析优化锰过氧化物酶培养基条件
Optimization of culture medium to produce manganese peroxidase by using Box-Behnken experimental design and response surface methodology
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摘要: 以白腐真菌为出发菌株,利用Design-Expert8.05软件设计,采用三水平部分因子分析初始发酵产酶培养基中10个因子,确定麸皮、酵母膏和KH2PO4为产锰过氧化物酶(Mnp)的显著影响因子,根据Box-Benhnken的中心组合实验设计及三因素三水平的响应面分析,通过二次多项回归模型进行方差分析和回归拟合,预测了最佳产酶培养基条件为:麸皮、酵母膏和KH2PO4的添加量分别为10.75、3.37、0.095g/L,最大Mnp酶活预测值为4.06U/mL。验证实验Mnp酶活为4.15U/mL,与预测值十分接近。优化后的酶活与优化前相比,Mnp酶活提高了58.4%。Abstract: White rot fungi preserved in our laboratory was used to produce manganese peroxidase.Design Expert 8.05 software was used to optimize the culture medium producing this peroxidase.3-Level factorial analysis was used to analyze the main effect factors from 10 factors.The result showed that the wheat bran, yeast extract and KH 2 PO 4 were the significant factors.According to Box-Behnken experimental design and response surface methodology, the second-order equation model was established by regression analysis of experimental data.The predicted optimal addition of wheat bran, yeast extract and KH 2 PO 4 were 10.75、3.37 and 0.095g/L respectively, and the predicted maximum manganese peroxidase could reach 4.06U/mL.The manganese peroxidase activity of verification experiment was 4.15U/mL, which was very close to the predicted value.The manganese peroxidase activity under the optimization condition increased by 58.4% compared with the original enzyme production medium.