产中性蛋白酶芽孢杆菌配伍发酵响应面法研究

Study on Fermentation Conditions of Neutral Protease from Bacillus Compatibility by Response Surface Methodology

目的:利用响应面法对芽孢杆菌配伍产中性蛋白酶的发酵培养基和发酵条件进行优化,确定最佳产酶条件。方法:通过Minitab软件的筛选试验设计对影响中性蛋白酶发酵的相关因素进行评价,用最陡爬坡试验逼近最大产酶区域,采用响应面分析法确定主要影响因素的最佳水平。结果:筛选出具有显著正效应的麸皮、接种量;具有显著负效应的装液量、温度。当麸皮的质量分数2.93%,温度32℃,接种量5.75%,装液量15 mL/50 mL时,酶活力的最大值为209.6867 U/mL。3次验证试验的平均值与预测值接近,相关系数为84.36%,酶活力提高了42%。结论:采用筛选试验、最陡爬坡试验及响应面分析法相结合,能够最大程度地缩短生产时间,降低开发成本,提高经济效益,有利于中性蛋白酶的工业化推广和应用。

Objective：Fermentation medium and conditions of neutral protease from Bacillus compatibility group were optimized by response surface analysis and the optimum condition of enzyme production was established.Method：Twolevel factorials of Plackett-Burman by Minitab software were constructed to evaluate the influence of related factors.The path of steepest ascent was used to approach the optimal region of the protease production.Then the optimized level of these factors were determined by response surface methodology.Result： The results showed that wheat bran and amount of inoculums influenced protease production positively while liquid volume and fermentation temperature influenced negatively,the highest anticipation protease activity is up to 209.6867 U/mL when the conditions are as follows： the concentration of wheat bran is 2.93%,incubation temperature is 32 ℃,amount of inoculums is 5.75% and liquid volume is 15 mL/50 mL.The results of three verification test approach the forecast value,enzymatic activity increased 42%,the correlation coefficient was 84.36%.Conclusion： Above these can shorten the production time,depress the developed cost,rise the economic returns,and easy to neutral protease industrialized extend and application.