响应面法优化海洋弧菌产褐藻胶裂解酶发酵培养基

Optimization of fermentation medium of alginate lyase produced by marine Vibrio sp. by response surface methodology

采用响应面法优化海洋弧菌X511的产酶发酵培养基提高其胞内褐藻胶裂解酶产量。通过单因素试验研究了不同碳源、不同氮源、海藻酸钠、氯化钠、蛋白胨、硫酸亚铁、硫酸镁和磷酸氢二钾对菌株产胞内酶活力的影响,在此基础上,利用Plackett-burman试验确定海藻酸钠、氯化钠、蛋白胨和硫酸镁对胞内酶产量的影响。通过响应面试验构建回归方程,结果表明,最佳发酵培养基成分为海藻酸钠9.0g/L,NaCl 31.6g/L,蛋白胨15.0g/L,MgSO_4·7H_2O 1.5g/L。在此条件下该菌株的胞内褐藻胶裂解酶的活力为(20.65±0.14)U/mL,较优化前的酶活提高了64.4%。

In order to improve the enzyme production of extracellular alginate lyase produced by marine Vibrio sp. X511, the fermentation medium was optimized by response surface methodology. The effects of different carbon sources, nitrogen sources, sodium alginate, NaCl, peptone, FeSO4, MgSO4·7H2O and K2HPO4 on the enzyme activity of the alginate lyase produced by the strain were studied by single factor experiment. On this basis, subsequently, the effect of sodium alginate, NaCl, MgSO4·7H2O and peptone on the intracellular enzyme production was determined by Plackett-Burman experiments. The regression equation was established by the response surface test, and the results showed that the optimum fermentation medium components were as followed： sodium alga acid 9.0 g/L, NaCl 31.6 g/L, peptone 15.0 g/L, MgSO4·7H2O 1.5 g/L. Under these conditions, the enzyme activity of the intracellular alginate lyase was （20.65±0.14） U/ml, which was 64.4% higher than before optimization.