Metabolic impact of redox cofactor perturbations in Saccharomyces cerevisiae |
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| Authors: | Jin Hou Nuno F. Lages Marco Oldiges Goutham N. Vemuri |
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| Affiliation: | 1. Center for Microbial Biotechnology, BioSys, Technical University of Denmark, Lyngby 2800, Denmark;2. State Key Laboratory of Microbial Technology, Shandong University, Shanda Nanlu 27, Jinan 250100, PR China;3. Centro de Química e Bioquímica, Departamento de Química e Bioquímica, Faculdade de Ciências da Universidade de Lisboa, Edifício C8, 1749-016 Lisboa, Portugal;4. Forschungszentrum Juelich, Institute of Biotechnology 2, Leo-Brandt-Str., D-52425 Juelich, Germany;1. Department of Biochemistry and Cell Biology, Rice University, Houston, TX 77005, USA;2. Department of Bioengineering, Rice University, Houston, TX 77005, USA;3. Department of Chemical and Biomolecular Engineering, Rice University, Houston, TX 77251, USA;1. State Key Laboratory of Food Science and Technology, Jiangnan University, Wuxi 214122, China;2. Key Laboratory of Industrial Biotechnology, Ministry of Education, Jiangnan University, Wuxi 214122, China;3. Laboratory of Food Microbial-Manufacturing Engineering, Jiangnan University, Wuxi 214122, China;1. Department of Biotechnology, The University of Tokyo, Yayoi 1-1-1, Bunkyo-ku, Tokyo 113-8657, Japan;2. Department of Biological Chemistry, College of Bioscience and Biotechnology, Chubu University, 1200 Matsumoto-cho, Kasugai, Aichi 487-8501, Japan;1. Department of Chemical Engineering, Pennsylvania State University, University Park, PA 16802, United States;2. Department of Biological Engineering, Pennsylvania State University, University Park, PA 16802, United States |
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| Abstract: | Redox cofactors play a pivotal role in coupling catabolism with anabolism and energy generation during metabolism. There exists a delicate balance in the intracellular level of these cofactors to ascertain an optimal metabolic output. Therefore, cofactors are emerging to be attractive targets to induce widespread changes in metabolism. We present a detailed analysis of the impact of perturbations in redox cofactors in the cytosol or mitochondria on glucose and energy metabolism in Saccharomyces cerevisiae to aid metabolic engineering decisions that involve cofactor engineering. We enhanced NADH oxidation by introducing NADH oxidase or alternative oxidase, its ATP-mediated conversion to NADPH using NADH kinase as well as the interconversion of NADH and NADPH independent of ATP by the soluble, non-proton-translocating bacterial transhydrogenase. Decreasing cytosolic NADH level lowered glycerol production, while decreasing mitochondrial NADH lowered ethanol production. However, when these reactions were coupled with NADPH production, the metabolic changes were more moderated. The direct consequence of these perturbations could be seen in the shift of the intracellular concentrations of the cofactors. The changes in product profile and intracellular metabolite levels were closely linked to the ATP requirement for biomass synthesis and the efficiency of oxidative phosphorylation, as estimated from a simple stoichiometric model. The results presented here will provide valuable insights for a quantitative understanding and prediction of cellular response to redox-based perturbations for metabolic engineering applications. |
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| Keywords: | Cofactor metabolism Redox metabolism Energy metabolism Saccharomyces cerevisiae Stoichiometric model |
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