重组大肠杆菌表达绿色气球菌丙酮酸氧化酶的条件优化  

Optimization of recombinant Escherichia coli cultivation for Aerococcus viridans pyruvate oxidase expression

作　　者：卢俊文[1] 张薷月 何宣贝 周紫薇 吕嘉猛 施沁怡 张建国[1] LU Junwen;ZHANG Ruyue;HE Xuanbei;ZHOU Ziwei;LYU Jiameng;SHI Qinyi;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2期1-6,共6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);微创励志创新基金项目

摘　　要：本文优化了表达绿色气球菌丙酮酸氧化酶的重组大肠杆菌的诱导条件。优化了诱导温度、诱导剂IPTG浓度、诱导时重组大肠杆菌的浓度、溶氧等条件后发现最佳的诱导条件为25℃、IPTG 0.5 mmol/L、OD_(600)0.5细胞浓度、250 m L摇瓶中50 m L发酵液。该条件下重组大肠杆菌诱导24 h后最终的丙酮酸氧化酶活力达到7 396.9 U/L,与优化前相比提高了4倍,是目前报道的最高水平。本研究结果为大肠杆菌大量表达有活性的丙酮酸氧化酶,以及丙酮酸氧化酶的生产应用奠定了基础。A recombinant Escherichia coli expressing Aerococcus viridans pyruvate oxidase gene(pod)was used for high level pyruvate oxidase through cultivation optimization.The optimal induction conditions were as follows:25℃,0.5 mmol/L IPTG,OD 600 0.5 initial cell density,50 mL working volume in 250 mL flask.After optimization under above conditions,the highest level of pyruvate oxidase was obtained as high as 7 396.9 U/L for 24 h induction,which increased 4-fold than that before optimization.This was the highest level of the current reported.This research provided a solid base for high level pyruvate oxidase production.

关 键 词：丙酮酸氧化酶 大肠杆菌 发酵优化 可溶性蛋白表达 

分 类 号：TQ925[轻工技术与工程—发酵工程]