Functional balance between enzymes in malonyl-CoA pathway for 3-hydroxypropionate biosynthesis |
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| Institution: | 1. CAS Key Lab of Biobased Materials, Qingdao Institute of Bioenergy and Bioprocess Technology, Chinese Academy of Sciences, Qingdao 266101, China;2. University of Chinese Academy of Sciences, Beijing 100049, China;3. Institute of Oceanology, Chinese Academy of Sciences, Qingdao 266071, China;3. Max-Planck-Institut for Biophysik, D-60438 Frankfurt am Main, Germany;4. Mikrobiologie Institut for Biologie II, Universität Freiburg, D-79104 Freiburg, Germany;5. Evonik Industries AG, D-45128 Essen, Germany;1. The Novo Nordisk Foundation Center for Biosustainability, Technical University of Denmark, Kogle Allé 6, 2970 Hørsholm, Denmark;2. Department of Biology and Biological Engineering, Chalmers University of Technology, Göteborg, Sweden;3. The Novo Nordisk Foundation Center for Biosustainability, Chalmers University of Technology, Göteborg, Sweden;1. Department of Chemical Engineering, Ulsan National Institute of Science and Technology (UNIST), Ulsan 44919, Republic of Korea;2. Department of Biomedical Engineering, Ulsan National Institute of Science and Technology (UNIST), Ulsan 44919, Republic of Korea;1. Laboratory of Synthetic Microbiology, School of Chemical Engineering & Technology, Tianjin University, Tianjin 300072, P.R. China;2. Key Laboratory of Systems Bioengineering, Ministry of Education of China, Tianjin 300072, P.R. China;3. Synbio Research Platform, Collaborative Innovation Center of Chemical Science and Engineering, Tianjin, P.R. China;1. Interdisciplinary Program of Bioengineering, Seoul National University, Seoul 151-921, South Korea;2. Department of Agricultural Biotechnology and Center for Food and Bioconvergence, Seoul National University, Seoul 151-921, South Korea;3. Department of Bio and Fermentation Convergence Technology, Kookmin University, Seoul 136-702, South Korea |
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| Abstract: | 3-Hydroxypropionate (3HP) is an important platform chemical, and four 3HP biosynthetic routes were reported, in which the malonyl-CoA pathway has some expected advantages but presented the lowest 3HP yield. Here, we demonstrated that this low yield was caused by a serious functional imbalance between MCR-C and MCR-N proteins, responsible for the two-step reduction of malonyl-CoA to 3HP. Then we minimized the enzyme activity imbalance by directed evolution of rate-limiting enzyme MCR-C and fine tuning of MCR-N expression level. Combined with culture conditions optimization, our engineering approaches increased the 3HP titer 270-fold, from 0.15 g/L to 40.6 g/L, representing the highest 3HP production via malonyl-CoA pathway so far. This study not only significantly improved the 3HP productivity of recombinant Escherichia coli strain, but also proved the importance of metabolic balance in a multistep biosynthetic pathway, which should be always considered in any metabolic engineering study. |
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| Keywords: | 3-hydroxypropionate Malonyl-CoA pathway Malonyl-CoA reductase Enzymatic activity imbalance |
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