响应面法优化紫红红球菌的发酵培养基

Optimization of the Fermentation Medium of Rhodococcus rhodochrous Using Response Surface Methodology

采用响应面法优化腈水合酶产生紫红红球菌的发酵培养基。以天冬酰胺、磷酸氢二钾、磷酸二氢钾、酵母抽提物、脲素、氯化钴为影响因子,以菌体干质量和酶活为响应值,进行多元二次响应回归分析,结果显示,最优培养基为:天冬酰胺含量0.04%、磷酸氢二钾0.05%、磷酸二氢钾0.03%、酵母抽提物(FM818)5.48%、脲素0.55%、氯化钴0.01%;在摇瓶中进行验证试验,优化条件下菌体干质量为33.98mg·mL-1、酶活为167.57U·mL-1;对照条件下菌体干质量26.6mg·mL-1、酶活为125.63U·mL-1,此优化培养基使干质量、酶活分别提高了27.74%和33.38%。

This research, response surface methodology（RSM） was utilized to optimize the fermentation medium of Rhodococcus rhodochrous which can produce nitrile hydratase. Asparagines,dipotassium hydrogen phosphate, potassium dihydrogen phosphate, yeast extract, urea, and cobalt chloride were selected as influencing factors . Dry cell weight（DCW） and enzyme activity was used as the response values and RSM was applied to analysis the results. The optimum fermentation medium obtained by RSM analysis were as follows,0.04% asparagine, 0.05% dipotassium hydrogen phosphate, 0.03% potassium dihydrogen phosphate, 5.48% yeast extract, 0.55% urea and 0.01% cobalt chloride. Finally, Verification test was accomplished in shaking flasks, DCW and enzyme activity were 33.98 mg·mL-1 and 167.57 U·mL-1 respectively, compared to that of the control （DCW and enzyme activity were 26.6 mg·mL-1 and 125.63 U·mL-1 respectively）, DCW and enzyme activity were increased by 27.74% and 33.38%, respectively.