响应面法优化醋酸钙不动杆菌菌株23的脂肪酶产酶条件

The Lipase Production Optimization of Acinetobacter calcoaceticus 23 Using Response Surface Methodology

关键词:  脂肪酶  醋酸钙不动杆菌  Plackett-Burman设计  响应面  优化

Key words:  lipase  Acinetobacter calcoaceticus  Plackett-Burman  response surface methodology  optimization

摘要:

采用响应面法对醋酸钙不动杆菌(Acinetobacter calcoaceticus)菌株23的发酵产脂肪酶培养基进行优化实验。实验首先考察该菌株产酶所需的最适碳源和氮源,然后用Plackett-Burman法设计实验确定影响产酶的主要影响因素,用最陡爬坡实验逼近最大响应区域,最后通过Box-Behnken方法进行二次回归分析得到最佳的产酶条件为:营养肉汤18.4%、蛋白胨9.18%、吐温800.52%、K₂HPO₄ 0.2%,橄榄油1%,MgSO₄ 0.05%,CaCl₂ 0.05%,FeSO₄ 0.1%。在该优化条件下,发酵液的脂肪酶活力可以达到15.2U/ml,比优化前提高3倍。

Abstract:

Response surface methodology (RSM)was employed to optimize culture medium for lipase production of Acinetobacter calcoaceticus 23. Firstly, the best carbon and nitrogen sources was investigated. Then a Plackett-Burman design was used to investigate the effects of different factor in the ferment medium, the 3 significtant is:broth, peptone, tween 80, and then steepest ascent procedures were employed to define optimal response region for these three factors. Finally, Box-Behnken was employed to design 17 experiments. The results of experiments was analyzed with Design-expert soft. The optimal lipase production medium is:18.4% nutrient broth, 9.18% tryptone, 0.52%, tween 80 0.2%, K₂HPO₄, 1%, olive oil 1%, MgSO₄ 0.05%, CaCl₂ 0.05%, FeSO₄ 0.1%, the yield of enzyme activity achieved 15.2 U/ml, the yield is increased by 3 times.