响应面法设计优化阿维菌素化学合成发酵培养基

Development and optimization of a fermentation chemically defined medium composition for the production of avermectins by response surface methodology

目的设计优化一种化学合成发酵培养基,为阿维菌素产生菌生理生化研究提供基础。方法在单因素实验的基础上运用响应面分析的方法,对9种因素进行Plackett-Burman设计筛选得到3个显著因子,并对其进行最陡爬坡试验和Box-Behnken试验,利用Design-Expert V8.06分析软件进行回归分析得到最优组合。结果优化后化学合成培养基的组成为:葡萄糖12g/L,麦芽糖18g/L,苏氨酸1.98g/L,50%乳酸钠0.5mL/L,K_2HPO_4·3H_2O 0.3g/L,MgSO_4·7H_2O 0.5g/L,NaCl 0.5g/L,FeSO_4·7H_2O0.01g/L,MnSO_4·H_2O 0.015g/L,MOPS 5g/L。结论经验证,该培养基的产素能力比目前的清亮培养基提高了397.4%,并具有透明清亮且产素稳定等特点,为以后阿维链霉菌的理论和生产研究奠定了基础。

Objective To design a chemically defined medium （CDM） for Streptomyces avermitilis which produces avermectins. Methods On the base of the single factor experiment, using the methodology of response surface analysis, three significant factors were obtained from nine influencing factors of Plackett-Burman experiments. In view of the three selected significant factors, the path of steepest ascent and the Box-Behnken design were used to approach the optimal region of the employed to get the best conditions. Results The optimal CDM medium was determined as fermentation conditions. The multiple regression analysis with Design-expert V8.06 was as follows： glucose 12g/L, maltose 18g/L, threonine 1.98g/L, 50% sodium lactate 0.5mL/L, KEHPO4·3H2O 0.3g/L, MgSO4·7H2O 0.Sg/L, NaCl 0.5g/L, FeSO4·7H2O 0.01g/L, MnSO4·H2O 0.015g/L, and MOPS 5g/L. Conclusion The production capacity of the CDM was 397.4% higher than that of the existing transparent medium, which has great potentiality for the future research on S. avermitilis.