响应面法优化表达丙酮酸氧化酶重组大肠杆菌培养基  

Optimization of recombinant Escherichia coli cultivation for pyruvate oxidase expression using response surface methodology

作　　者：姚兵莉 卢俊文[1] 张建国[1] YAO Bingli;LU Junwen;ZHANG Jianguo(Institute of Food Science and Engineering, University of Shanghai for Science and Technology, Shanghai 200093)

机构地区：[1]上海理工大学食品科学与工程研究所,上海200093

出　　处：《工业微生物》2018年第6期25-31,共7页Industrial Microbiology

基　　金：国家自然科学基金(No.21306112);上海市自然科学基金资助项目(No.13ZR1429100)

摘　　要：本研究利用响应面法优化重组大肠杆菌的培养基提高了其表达丙酮酸氧化酶的产量。首先,单因素优化实验表明pH、甘油浓度、酵母粉浓度、胰蛋白胨浓度分别为pH 7、0. 3%、1. 2%、0. 6%时有利于重组大肠杆菌表达丙酮酸氧化酶。进而,利用Box-Behnken法对甘油、酵母粉、胰蛋白胨设计三因素三水平实验,通过响应面回归分析,得到模型预测的最优培养基条件。在0. 6%甘油、3. 18%酵母粉和0. 44%胰蛋白胨的培养基条件下重组大肠杆菌产丙酮酸氧化酶有最高活力。验证试验结果丙酮酸氧化酶活力达到(17 194. 9±504. 9) U/L,与模型的预测值相近,比未优化前提高了1. 7倍。本研究结果为丙酮酸氧化酶的大规模生产和工业化应用提供了基础。Pyruvate oxidase is an important biochemical agent for analytical kits.The medium of a recombinant Escherichia coli expressing Aerococcus viridans pyruvate oxidase gene(pod)was optimized using response surface methodology.First,the optimized results of single parameter experiments were obtained as follows:pH 7.0,glycerol 0.3%,yeast extract 1.2%and peptone 0.6%,respectively.Then,a three factors-three levels experiment was carried out based on the Box-Behnken design.Under the optimized conditions of 0.6%glycerol,3.18%yeast extract,0.44%peptone,the highest pyruvate oxidase activity was(17 194.9±504.9)U/L in the validation experiment.It was similar to the predicted value of the model,which was 1.7 times higher than that before optimization.The results of this study provided a basis for large-scale production and industrial application of pyruvate oxidase.

关 键 词：丙酮酸氧化酶 大肠杆菌发酵 培养基优化 响应面 

分 类 号：TQ925[轻工技术与工程—发酵工程]