响应面法优化紫红红球菌发酵培养基.docVIP

响应面法优化紫红红球菌发酵培养基 　　摘 要：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33.98 mg?mL-1、酶活为167.57 U?mL-1；对照条件下菌体干质量26.6 mg?mL-1、酶活为125.63 U?mL-1，此优化培养基使干质量、酶活分别提高了27.74%和33.38%。 　　关键词：响应面分析；紫红红球菌；腈水合酶；丙烯酰胺 　　中图分类号：TQ225.1 文献标识码：A DOI编码：10.3969/j.issn.1006－6500.2011.05.005 　　 　　Optimization of the Fermentation Medium of Rhodococcus rhodochrous Using Response Surface Methodology 　　LI Xiao1,2,3, ZHANG Ya2 , LI Jian-hua2, YAO Juan1, LI Pei1, LIANG Yun-xiang3, LI Zhi-hong1, YU Xue-feng1 　　(1.Angel Yeast Company Limited,Yichang,Hubei 443003,China; 2.College of Chemistry and Biology, Three Gorges Univerity,Yichang,Hubei 443002,China; 3.College of Life Science, Huazhong Agricultural University,Wuhan, Hubei 430068,China) 　　Ａｂｓｔｒａｃｔ: 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. 　　Key words: response surface methodology; Rhodococcus rhodochrous; nitrile hydratase; acrylamide 　　 　　丙烯酰胺(简称AM，结构式为CH=CH―CONH2)是