Quantitative analysis and engineering of fatty acid biosynthesis in E. coli |
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| Authors: | Tiangang Liu Harmit Vora Chaitan Khosla |
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| Institution: | 1. Department of Chemistry, Stanford University, Stanford, CA 94305-5025, USA;2. Department of Chemical Engineering, Stanford University, Stanford, CA 94305, USA;3. Department of Biochemistry, Stanford University, Stanford, CA 94305, USA;1. Department of Chemical and Biological Engineering, Iowa State University, Ames, IA 50011, USA;2. Chemical Engineering, School for Engineering of Matter, Transport, and Energy, Arizona State University, Tempe, AZ 85287-6106, USA;1. Department of Chemical and Biological Engineering, University of Wisconsin-Madison, Madison, WI, United States;2. Microbiology Doctoral Training Program, University of Wisconsin-Madison, Madison, WI, United States;3. Department of Biology and Biological Engineering, Chalmers University of Technology, Göteborg, Sweden;1. Key Laboratory of Combinatorial Biosynthesis and Drug Discovery, Ministry of Education and Wuhan University School of Pharmaceutical Sciences, Wuhan 430071, PR China;2. State Key Laboratory of Microbial Metabolism, School of Life Sciences and Biotechnology, Shanghai Jiao Tong University, Shanghai 200240, PR China;3. Hubei Engineering Laboratory for Synthetic Microbiology, Wuhan Institute of Biotechnology, Wuhan 430075, PR China;1. College of Engineering, University of Georgia, Athens, GA 30602, USA;2. Department of Chemistry and Chemical Biology, Center for Biotechnology and Interdisciplinary Studies, Rensselaer Polytechnic Institute, Troy, NY 12180, USA;3. BioChemical Engineering Program, College of Engineering, University of Georgia, Athens, GA 30602, USA;1. College of Food Science and Technology, Nanjing Agricultural University, Nanjing, Jiangsu 210095, China;2. Department of Food Science and Technology, College of Light Industry and Food Engineering, Nanjing Forestry University, Nanjing 210037, China |
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| Abstract: | Fatty acids are central hydrocarbon intermediates in the biosynthesis of diesel from renewable sources. We have engineered an Escherichia coli cell line that produces 4.5 g/L/day total fatty acid in a fed-batch fermentation. However, further enhancement of fatty acid biosynthesis in this cell line proved unpredictable. To develop a more reliable engineering strategy, a cell-free system was developed that enabled direct, quantitative investigation of fatty acid biosynthesis and its regulation in E. coli. Using this system, the strong dependence of fatty acid synthesis on malonyl-CoA availability and several important phenomena in fatty acid synthesis were verified. Results from this cell-free system were confirmed via the generation and analysis of metabolically engineered strains of E. coli. Our quantitative findings highlight the enormous catalytic potential of the E. coli fatty acid biosynthetic pathway, and target specific steps for protein and metabolic engineering to enhance the catalytic conversion of glucose into biodiesel. |
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