Penicillium sp. 1523产柚苷酶摇瓶发酵培养基优化

Optimization of Shaking-flask Fermentation Medium for Naringinase Production by Penicillium sp. 1523

采用单因素试验、Plackett-Burman设计和响应面分析相结合的方法,对Penicillium sp.1523产柚苷酶的摇瓶发酵培养基配方进行优化。单因素试验结果显示:发酵培养基中的最优碳源为玉米粉,最优氮源为豆饼粉;Plackett-Burman设计筛选出影响柚苷酶产量的3个重要因素为玉米粉、豆饼粉和柚苷,在此基础上运用最陡爬坡试验逼近最大响应值区域,再利用Box-Behnken试验设计及响应面分析法进行回归分析,获得最佳培养基配方为:玉米粉31.14g/L、豆饼粉31.53g/L、柚苷1.65g/L、K2HPO4 1.00g/L、ZnSO4 0.10g/L、MgSO4.7H2O 0.06g/L、CaCl20.10g/L。在优化后的条件下摇瓶发酵产柚苷酶酶活力为(891.79±6.33)U/mL,与模型预测值接近,发酵产酶量比优化前提高70.8%。

Single-factor method,Plackett-Burman design and response surface methodology were combinedly used to optimize the fermentation medium composition for naringinase production by Penicillium sp.1523 at shake flask level.The optimum carbon and nitrogen sources for the growth of Penicillium sp.1523 were corn meal and bean cake powder,respectively.Using Plackett-Burman design,corn meal,bean cake powder and naringin were screened out of 7 factors as main affecting variables of naringinase production and steepest ascent method was then used to approach their maximum response regions,followed by central composite design,multiple regression analysis and response surface analysis.The optimum medium formula（g/L） determined was composed of corn meal 31.14,bean cake powder 31.53,naringin 1.65,dipotassium hydrogen phosphate 1.00,zinc sulfate 0.10,heptahydrate magnesium sulfate 0.06 and calcium chloride 0.10,and the resulting naringinase activity in fermentation broth reached up to（891.79 ± 6.33） U/mL and was 70.8% higher than before the optimization and consistent with the maximum predicted value.