文献类型：期刊文献

英文题名：Significantly enhanced production of recombinant nitrilase by optimization of culture conditions and glycerol feeding

作者：Liu, Jun-Feng[1];Zhang, Zhi-Jun[1];Li, Ai-Tao[1];Pan, Jiang[1];Xu, Jian-He[1]

机构：[1]E China Univ Sci & Technol, State Key Lab Bioreactor Engn, Shanghai 200237, Peoples R China

年份：2011

卷号：89

期号：3

起止页码：665

外文期刊名：APPLIED MICROBIOLOGY AND BIOTECHNOLOGY

收录：;EI(收录号：20110513633624);WOS:【SCI-EXPANDED(收录号:WOS:000286429400019)】；

基金：This work was financially supported by the National Natural Science Foundation of China (Nos. 20672037, 20773038, and 20902023), Ministry of Science and Technology, PR China (Nos. 2009ZX09501-016 and 2009CB724706), China National Special Fund for State Key Laboratory of Bioreactor Engineering (No. 2060204), and Shanghai Municipal Fund for Natural Science (No. 07ZR14030).

语种：英文

外文关键词：Nitrilase; Optimization; Fermentation; Glycerol feeding; Recombinant Escherichia coli

摘要：The production of a recombinant nitrilase expressed in Escherichia coli JM109/pNLE was optimized in the present work. Various culture conditions and process parameters, including medium composition, inducer, induction condition, pH and temperature, were systematically examined. The results showed that nitrilase production in E. coli JM109/pNLE was greatly affected by the pH condition and the temperature in batch culture, and the highest nitrilase production was obtained when the fermentation was carried out at 37A degrees C, initial pH 7.0 without control and E. coli was induced with 0.2 mM isopropyl-beta-d-thiogalactoside at 4.0 h. Furthermore, enzyme production could be significantly enhanced by adopting the glycerol feeding strategy with lower flow rate. The enzyme expression was also authenticated by sodium dodecyl phosphate polyacrylamide gel electrophoresis analysis. Finally, under the optimized conditions for fed-batch culture, cell growth, specific activity and nitrilase production of the recombinant E. coli were increased by 9.0-, 5.5-, and 50-fold, respectively.