生防菌菌株A的摇瓶培养条件及发酵工艺

[目的]优化生防菌菌株A的培养条件与液体发酵工艺。[方法]采用均匀试验设计方法、回归分析方法和分批补料发酵工艺，研究生防菌菌株A的培养条件与液体发酵工艺的优化。[结果]结果表明，适合菌株A摇瓶培养的最佳条件为：培养温度35℃，摇瓶装量12％，接种量3．0％，初始pH值7．2。20L发酵罐分批补料发酵参数为：种子培养基为YDC培养基，培养时间24h，接种量3．0％；发酵培养基：葡萄糖1．0％，酵母膏1．0％，CaCO3 1．0％，pH值7．2；流加培养基：葡萄糖1．5％，酵母膏0．5％，制成混合液一起流加，发酵8h开始流加；发酵周期（34±2）h，在以上发酵条件下，发酵液芽孢浓度达（3．410±0．151）×10^9cfu／ml。[结论]该试验条件下所获得发酵液芽孢数浓度较高，可用于进一步制备生防菌剂。

Studies on the Culture Condition and Fermentation Crafts of A Strain of Bacillus subtilis

[Objective]The purpose was to optimize the culture conditions and fermentation crafts of strain A,a kind of Bacillus subtilis(CGMCC No.1982),and in order to obtain high spores concentration of fermentation broth and establish scientific foundation for empoldering the strain A.[Method] Uniform design tests,regression analysis method and fed-batch fermentation techniques were used.[Result] The results showed that the optimum conditions for shaking flask culture of strain A were as follows: 35 ℃ of culture temperature,culture broth quantity 12%,inoculation amount 3.0%,initial pH value 7.2.The fed-batch fermentation crafts were ascertained by 20 L fermenter.The seed was prepared by using YDC as seed culture medium(glucose 20 g,yeast extracts 10 g,CaCO3 20 g and water 1 000 ml),culture time 24 h and inoculation amount 3.0%.The fermentation medium(glucose 1.0%,yeast extracts 1.0%,CaCO31.0% and pH value 7.2) and the feeding medium,a mixture of glucose 1.5% and yeast extracts 0.5%,were confirmed.The feeding began at 8 h later and the fermenting period lasted for(34±2) h,the spore concentration could reach to(3.410±0.151) ×109 cfu/ml.[Conclusion]The culture conditions and fermentation crafts of strain A to produce high concentration of spores was validated,which will establish scientific foundation for empoldering the strain A.