响应面法优化枯草芽孢杆菌BS-1101原生质体制备条件

Optimization of Protoplast Formation Conditions of Bacillus Subtilis BS-1101 by Using Response Surface Methodology

实验以枯草芽孢杆菌BS-1101原生质体的制备率为目的,通过中心组合试验对影响枯草芽孢杆菌BS-1101原生质体的制备率的三个因素:酶解温度、酶解时间、酶浓度进行了研究评价,通过中心组合试验设计与响应面分析方法得到二次回归模型程,由回归方程得到,在酶解温度37.27℃、酶浓度为5.152 mg.mL-1、酶解时间88.8 min的条件下得到枯草芽孢杆菌BS-1101制备率为77.02%,与方程预测值为较接近。结果表明利用响应面法能够很好地对枯草芽孢杆菌原生质体制备条件进行优化。对于枯草芽孢杆菌原生质体制备条件的优化有一定参考价值。

The central composite design combined with response surface methodology was employed to optimize the protoplast formation conditions of Bacillus subtilis BS-1101.The critical parameters,such as lysozyme concentration,temperature,and contact time which affects the formation of protoplast of the Bacillus subtilis BS-1101 were studied using a novel technique named response surface methodology.Second-order multiple regression analysis was used to build a predicted model of the combined effects of independent variables（enzyme concentration,temperature and effect time） for maximum the yield of protoplast of Bacillus subtilis BS-1101.The optimal parameters to obtain the highest protoplast yield were as followed： temperature was 37.27 ℃,effect time was 88.8min,enzyme concentration was 5.152 mg·mL-1.The average experimental protoplast yield under the optimum conditions was found to be 77.02%,which agreed with the predicted value.The proposed method showed high degree of reproducibility.The central composite design was employed to optimize the protoplast formation conditions of Bacillus subtilis BS-1101.