Engineering Escherichia coli for production of C12–C14 polyhydroxyalkanoate from glucose |
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| Authors: | Daniel E Agnew Amanda K Stevermer JTyler Youngquist Brian F Pfleger |
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| Institution: | 1. Department of Chemical and Biological Engineering, University of Wisconsin-Madison, Madison, WI 53706, USA;2. Great Lakes Bioenergy Research Center, University of Wisconsin-Madison, Madison, WI 53706, USA;1. Department of Innovative and Engineered Materials, Tokyo Institute of Technology, 4259 Nagatsuta, Midori-ku, Yokohama 226-8502, Japan;2. Department of Materials Science and Chemical Engineering, Kitakyushu National College of Technology, 5-20-1 Shii, Kokuraminami-ku, Kitakyushu 802-0985, Japan;3. Faculty of Environmental and Symbiotic Sciences, Prefectural University of Kumamoto, 3-1-100 Tsukide, Higashi-ku, Kumamoto 862-8502, Japan;1. School of Biological Sciences, Universiti Sains Malaysia, 11800 Penang, Malaysia;2. Faculty of Science, Department of Biochemistry, Bauchi State University, Gadau, Bauchi, Nigeria;1. Huck Institutes of Life Sciences, Pennsylvania State University, University Park, PA, USA;2. Department of Chemical and Biological Engineering, Iowa State University, Ames, IA, USA;3. Department of Chemical Engineering, Pennsylvania State University, University Park, PA, USA;1. Department of Laboratory Medicine and Biotechnology, Tzu Chi University, College of Medicine, Hualien, Taiwan, ROC;2. Graduate School of Biotechnology and Bioengineering, Yuan Ze University, Chung-Li, Taiwan, ROC;3. Department of Chemical Engineering and Materials Science, Yuan Ze University, Chung-Li, Taiwan, ROC |
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| Abstract: | Demand for sustainable materials motivates the development of microorganisms capable of synthesizing products from renewable substrates. A challenge to commercial production of polyhydroxyalkanoates (PHA), microbially derived polyesters, is engineering metabolic pathways to produce a polymer with the desired monomer composition from an unrelated and renewable source. Here, we demonstrate a metabolic pathway for converting glucose into medium-chain-length (mcl)-PHA composed primarily of 3-hydroxydodecanoate monomers. This pathway combines fatty acid biosynthesis, an acyl-ACP thioesterase to generate desired C12 and C14 fatty acids, β-oxidation for conversion of fatty acids to (R)-3-hydroxyacyl-CoAs, and a PHA polymerase. A key finding is that Escherichia coli expresses multiple copies of enzymes involved in β-oxidation under aerobic conditions. To produce polyhydroxydodecanoate, an acyl-ACP thioesterase (BTE), an enoyl-CoA hydratase (phaJ3), and mcl-PHA polymerase (phaC2) were overexpressed in E. coli ΔfadRABIJ. Yields were improved through expression of an acyl-CoA synthetase resulting in production over 15% CDW – the highest reported production of mcl-PHA of a defined composition from an unrelated carbon source. |
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