响应面法优化醋酸钙不动杆菌菌株23的脂肪酶产酶条件

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%、K2HPO40.2%,橄榄油1%,MgSO40.05%,CaCl20.05%,FeSO40.1%。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。

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

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.