文献类型：期刊文献

英文题名：Increasing glycolytic flux in Torulopsis glabrata by redirecting ATP production from oxidative phosphorylation to substrate-level phosphorylation

作者：Liu, L.M.[1]; Li, Y.[1,2,3]; Du, G.C.[1]; Chen, J.[1,4]

机构：[1] Key Laboratory of Industrial Biotechnology, School of Biotechnology, Southern Yangtze University, Wuxi, China; [2] School of Biotechnology, Southern Yangtze University, Wuxi, China; [3] State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology, Shanghai, China; [4] School of Biotechnology, Southern Yangtze University, 170 Huihe Road, Wuxi 214036, China

年份：2006

卷号：100

期号：5

起止页码：1043

外文期刊名：Journal of Applied Microbiology

收录：EI(收录号：20231713966388)

语种：英文

外文关键词：Antibiotics - Enzyme activity - Glucose - Substrates

摘要：Aims: This study aimed at further increasing the pyruvate productivity of a multi-vitamin auxotrophic yeast Torulopsis glabrata by redirecting ATP production from oxidative phosphorylation to substrate-level phosphorylation. Methods and Results: We examined two strategies to decrease the activity of F0F1-ATPase. The strategies were to inhibit F0F1-ATPase activity by addition of oligomycin, or to disrupt F0F1-ATPase by screening neomycin-resistant mutant. The addition of 0.05 mmol l-1 oligomycin to the culture broth of T. glabrata CCTCC M202019 resulted in a significantly decreased intracellular ATP level (35.7%) and a significantly increased glucose consumption rate (49.7%). A neomycin-resistant mutant N07 was screened and selected after nitrosoguanidine mutagenesis of the parent strain T. glabrata CCTCC M202019. Compared with the parent strain, the F0F1-ATPase activity of the mutant N07 decreased about 65%. As a consequence, intracellular ATP level of the mutant N07 decreased by 24%, which resulted in a decreased growth rate and growth yield. As expected, glucose consumption rate and pyruvate productivity of the mutant N07 increased by 34% and 42.9%, respectively. Consistently, the activities of key glycolytic enzymes of the mutant N07, including phosphofructokinase, pyruvate kinase and glyceraldyde-3-phosphate dehydrogenase, increased by 63.7%, 28.8% and 14.4%, respectively. In addition, activities of the key enzymes involved in electron transfer chain of the mutant N07 also increased. Conclusions: Impaired oxidative phosphorylation in T. glabrata leads to a decreased intracellular ATP production, thereby increasing the glycolytic flux. Significance and Impact of the Study: The strategy of redirecting ATP production from oxidative phosphorylation to substrate-level phosphorylation provides an alternative approach to enhance the glycolytic flux in eukaryotic micro-organisms. ? 2006 The Society for Applied Microbiology.