Engineering central metabolic modules of Escherichia coli for improving β-carotene production |
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| Institution: | 1. Tianjin Institute of Industrial Biotechnology, Chinese Academy of Sciences, Tianjin 300308, China;2. Key Laboratory of Systems Microbial Biotechnology, Chinese Academy of Sciences, Tianjin 300308, China;3. College of Biotechnology, Tianjin University of Science & Technology, Tianjin 300308, China;1. Department of Life Sciences, Pohang University of Science and Technology, 77 Cheongam-ro, Nam-gu, Pohang, Gyeongbuk 37673, Korea;2. Department of Chemical Engineering, Pohang University of Science and Technology, 77 Cheongam-ro, Nam-gu, Pohang, Gyeongbuk 37673, Korea;3. School of Interdisciplinary Bioscience and Bioengineering, Pohang University of Science and Technology, 77 Cheongam-ro, Nam-gu, Pohang, Gyeongbuk 37673, Korea;4. Department of Chemical and Biological Engineering, Korea University, 145 Anam-ro, Seongbuk-gu, Seoul 02841, Korea;1. Tianjin Institute of Industrial Biotechnology, Chinese of Academy of Sciences, Tianjin 300308, China;2. Key Laboratory of Systems Microbial Biotechnology, Tianjin 300308, China;3. College of Biotechnology, Tianjin University of Science and Technology, Tianjin 300457, China;1. Faculty of Life Science and Food Engineering, HuaiYin Institute of Technology, Huaian 223003, China;2. Division of Applied Life Science (BK21 Plus), PMBBRC, Gyeongsang National University, Jinju 52828, South Korea;3. Industrial Biotechnology Research Center, KRIBB, Daejeon 28116, South Korea;1. Institute of Bioengineering, College of Chemical and Biological Engineering, Zhejiang University, Hangzhou 310027, PR China;2. Key Laboratory of Biomass Chemical Engineering of Ministry of Education, Zhejiang University, Hangzhou 310027, PR China;1. Key Laboratory of Systems Microbial Biotechnology, Tianjin Institute of Industrial Biotechnology, Chinese Academy of Sciences, China;2. University of Chinese Academy of Sciences, China;3. College of Biotechnology, Tianjin University of Science & Technology, China |
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| Abstract: | ATP and NADPH are two important cofactors for production of terpenoids compounds. Here we have constructed and optimized β-carotene synthetic pathway in Escherichia coli, followed by engineering central metabolic modules to increase ATP and NADPH supplies for improving β-carotene production. The whole β-carotene synthetic pathway was divided into five modules. Engineering MEP module resulted in 3.5-fold increase of β-carotene yield, while engineering β-carotene synthesis module resulted in another 3.4-fold increase. The best β-carotene yield increased 21%, 17% and 39% after modulating single gene of ATP synthesis, pentose phosphate and TCA modules, respectively. Combined engineering of TCA and PPP modules had a synergistic effect on improving β-carotene yield, leading to 64% increase of β-carotene yield over a high producing parental strain. Fed-batch fermentation of the best strain CAR005 was performed, which produced 2.1 g/L β-carotene with a yield of 60 mg/g DCW. |
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