Plackett-Burman试验联用响应面法优化枯草芽孢杆菌产α-淀粉酶培养基营养成分

Optimization of Nutritional Components of Culture Medium forα-Amylase Produced by Bacillus subtilis Using Plackett-Burman

以枯草芽孢杆菌发酵液产α-淀粉酶为对象,考察发酵液营养组成成分条件。在发酵培养基组成单因素试验的基础上,结合Plackett-Burman筛选试验、营养组分最低添加量、最陡爬坡试验和响应面Box-Benhnken试验方法,选择出对试验结果具有显著影响的因素,建立各显著性因素的二次回归方程模型,根据试验实际情况,得到最佳发酵营养成分配方:糖蜜25.0 g/L、酵母粉7.6 g/L、硫酸锰6.0 g/L、磷酸二氢钾7.0 g/L、胰蛋白胨8.0 g/L、氯化钠3 g/L、磷酸二氢钠5 g/L、磷酸氢二钾1 g/L,平均发酵液淀粉酶酶活为(2084±21)U/g。调整后实测值与二次回归方程预测值吻合良好。

The production ofα-amylase by Bacillus subtilis fermentation broth is studied,and the nutritional components of fermentation broth are investigated.On the basis of single factor test of fermentation medium composition,the factors that have significant influence on the test results are selected and the quadratic regression equation model of each significant factor is established with Plackett-Burman screening test,the minimum additive amount of nutritional components and the steepest climbing test and response surface Box-Benhnken test method.According to the actual situatio n of the test,the optimal formula of fermentation nutritional components is obtained:molasses is 25.0 g/L,yeast powder is 7.6 g/L,manganese sulfate is 6.0 g/L,potassium dihydrogen phosphate is 7.0 g/L,tryptone is 8.0 g/L,sodium chloride is 3 g/L,sodium dihydrogen phosphate is 5 g/L,potassium dihydrogen phosphate is 1 g/L,the average amylase activity of fermentation broth is(2084±21)U/g.The measured value is in good agreement with the predicted value of quadratic regression equation.