响应面法优化枯草芽孢杆菌NHS1产芽孢发酵培养  

Optimization of fermentation medium composition by response surface methodology for the spore production of Bacillus subtilis

作　　者：胡瑞萍 丁贤[1,2] 李来好 李俊伟[1] 伍文超[2] 徐宁 

机构地区：[1]中国水产科学研究院南海水产研究所农业部水产品加工重点实验室,广州510300 [2]广州市海威水产科技有限公司,广州510300

出　　处：《生态学杂志》2018年第2期605-612,共8页Chinese Journal of Ecology

基　　金：国家自然科学基金项目(41606137);广东省自然科学基金项目(2015A030313855;2015A030310260);广州市科技计划项目(201707010439);农业部水产品质量加工重点实验室(NYJG201308)资助

摘　　要：为提高枯草芽孢杆菌(Bacillus subtilis)NHS1菌株发酵液中芽孢含量,采用单因素试验与响应曲面法优化试验相结合的方式,对该菌发酵培养基中的碳源、氮源以及无机盐等组成成分进行了优化。通过单因素试验、Plackett-Burman试验、最陡爬坡试验以及BoxBchnken试验构建响应方程,得到方程为:Y=-3.30+3.853X1+0.0928X2+0.623X3-0.913X1×X1-0.00704X2×X2-0.0433X3×X3-0.0033X1×X2-0.0700X1×X3+0.00167X2×X3。利用该方程预测得到最优培养基:淀粉1.88 g·L^(-1)、Na Cl 6.83 g·L^(-1)、玉米粉5.60 g·L^(-1),酵母粉10g·L^(-1),蛋白胨10 g·L^(-1),牛肉膏15 g·L^(-1),葡萄糖2 g·L^(-1),Mg SO43 g·L^(-1)。利用优化培养基,在36℃、170 r·min^(-1)条件下摇瓶发酵72 h,芽孢数达到2.42×109cfu·m L^(-1),比优化前提高1.5倍。Both one-factor-at-a-time method and response surface methodology were used to enhance the spore content of Bacillus subtilis NHS1 in fermentation broth. A response surface regression equation was established based on the results of one-factor-at-a-time, Plackett- Burman, steepest ascent and Box-Behnken experimental designs. The optimal medium formulation, as predicted from the equation, consisted of 1.88 g · L- starch, 6.83 g · L-1 NaCI, 5.60 g · L-1 corn flour, 10 g · L-1 yeast extract powder, 10 g · L-1 peptone, 15 g · L-1 beef extract, 2 g · L-1 glucose, 3 g · L-1 MgSO4. The yield of the spore reached 2.42×109cfu · mL-1 which was 1.5-fold higher than that before the optimization by using the optimized medium at 36 ℃ with shaking at a speed of 170 r ·min-1 for 72 h.

关 键 词：响应面分析法 枯草芽孢杆菌 产孢率 发酵培养基 

分 类 号：S963[农业科学—水产养殖]