多杀菌素种子培养基及发酵培养基的优化

摘要

Objective Saccharopolyspora spinosa CB11 was used as spinosad producing strain. Taking biomass and spinosad production in the fermentation medium as target parameters, carbon sources and nitrogen sources in the seed medium was optimized by single-factor test and orthogonal experimental design.Then fermentation medium was optimized by RSA, the significant factors was found and its optimal concentration was obtained. Methods Single-factor test, orthogonal experimental design and RSA. Results The optimal seed medium was obtained as following: glucose 10g/L, glycerol 5g/L, TSB 25g/L, corn steep liquor 10g/L, cotton seed protein 25g/L. When S. Spinosa CB11 grown in the optimzed seed medium was transferred into the fermentation medium, the spinosad production reached 411.26mg/L, increasing by 86.68% compared with original level. Then fermentation medium was optimized by RSA, three most significant factor was glucose, corn steep liquor and K2HPO4, its optimal concentration was respectively 66.6, 14.6 and 2.5g/L, the spinosad production reached 544.60g/L, increasing 24.48% compared with level before rnoptimization. Conclusion Through the optimization of seed medium and fermentation medium in spinosad fermentation,the most significant factor and its optimal concentration was obtained. Utilizing the optimized medium spinosad yield increased by 86.68%,which is a satisfactory result.%目的 以刺糖多孢菌CB11为试验菌株,以发酵培养基菌体浓度和多杀菌素产量为指标,通过单因素实验和正交试验对多杀菌素发酵的种子培养基的碳氮源成分进行优化,在此基础上通过响应面分析试验优化发酵培养基,找出最显著因素并确定其最佳值.方法 种子培养基碳氮源成分单因素实验和正交试验,发酵培养基响应面试验.结果 得到最优种子培养基配方为:葡萄糖10g/L,甘油5g/L,TSB25g/L,玉米浆10g/L,棉籽蛋白25g/L.在此培养基上生长的种子接入发酵培养基中所获得的多杀菌素产量达到411.26mg/L,较优化前水平(220.30mg/L)提高了86.68％.在此基础上,通过响应面分析试验优化了发酵培养基,得到对多杀菌素生产影响最显著的三个因素及最佳含最分别为葡萄糖66.6g/L、玉米浆14.6g/L、K2HPO4 2.5g/L,经验证获得的多杀菌素产量为544.60 g/L,较优化前产量提高了24.48％.结论 通过对多杀菌素发酵摇瓶种子培养基及发酵培养基的优化,找出了影响多杀菌素发酵的最显著因子及其最适含量,利用优化后的培养基发酵多杀菌素产量提高了86.68％,取得了较好的效果.