文献类型：期刊文献

英文题名：Combined 13C-assisted metabolomics and metabolic flux analysis reveals the impacts of glutamate on the central metabolism of high β-galactosidase-producing Pichia pastoris

作者：Liu, Ping[1]; Huang, Mingzhi[1]; Guo, Menglei[1]; Qian, Jiangchao[1]; Lin, Weilu[1]; Chu, Ju[1]; Zhuang, Yingping[1]; Zhang, Siliang[1]

机构：[1] State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology, No.130, Meilong Road, Shanghai, 200237, China

年份：2016

卷号：3

期号：1

外文期刊名：Bioresources and Bioprocessing

收录：EI(收录号：20224513087567)

语种：英文

外文关键词：Gene expression - Physiology - Recombinant proteins - Yeast

摘要：Background: Pichia pastoris is a popular recombinant protein expression system for its accessibility of efficient gene manipulation and high protein production. Sufficient supply of precursors, energy, and redox cofactors is crucial for high recombinant protein production. In our present work, we found that the addition of glutamate improved the recombinant β-galactosidase (β-gal) production by P. pastoris G1HL. Methods: To elucidate the impacts of glutamate on the central metabolism in detail, a combined 13C-assisted metabolomics and 13C metabolic flux analysis was conducted based on LC–MS/MS and GC–MS data. Results: The pool sizes of intracellular amino acids were obviously higher on glucose/glutamate (Glc/Glu). The fluxes in EMP entry reaction and in downstream TCA cycle were 50 and 67% higher on Glc/Glu than on Glc, respectively. While the fluxes in upstream TCA cycle kept almost unaltered, the fluxes in PPP oxidative branch decreased. Conclusion: The addition of glutamate leads to a remarkable change on the central metabolism of high β-galactosidase-producing P. pastoris G1HL. To meet the increased demands of redox cofactors and energy for higher β-galactosidase production on Glc/Glu, P. pastoris G1HL redistributes the fluxes in central metabolism through the inhibitions and/or activation of the enzymes in key nodes together with the energy and redox status. ? 2016, The Author(s).