Biosynthesis of poly(2-hydroxybutyrate-co-lactate) in metabolically engineered <Emphasis Type="Italic">Escherichia coli</Emphasis> |
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Authors: | Cheol Gi Chae You Jin Kim Se Jin Lee Young Hoon Oh Jung Eun Yang Jeong Chan Joo Kyoung Hee Kang Young-Ah Jang Hyuk Lee A-Reum Park Bong Keun Song Sang Yup Lee Si Jae Park |
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Institution: | 1.Department of Environmental Engineering and Energy,Myongji University,Yongin,Korea;2.Center for Bio-based Chemistry, Division of Convergence Chemistry,Korea Research Institute of Chemical Technology,Daejeon,Korea;3.Metabolic and Biomolecular Engineering National Research Laboratory, Department of Chemical and Biomolecular Engineering (BK21 Program),Center for Systems and Synthetic Biotechnology, and Institute for the BioCentury, KAIST,Daejeon,Korea;4.Division of Drug Discovery Research,Korea Research Institute of Chemical Technology,Daejeon,Korea |
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Abstract: | We have previously reported in vivo biosynthesis of polyhydroxyalkanoates containing 2-hydroxyacid monomers such as lactate and 2-hydroxybutyrate in recombinant Escherichia coli strains by the expression of evolved Clostridium propionicum propionyl-CoA transferase (PctCp) and Pseudomonas sp. MBEL 6-19 polyhydroxyalkanoate (PHA) synthase 1 (PhaC1 Ps6-19). Here, we report the biosynthesis of poly(2-hydroxybutyrate-co-lactate)P(2HB-co-LA)] by direct fermentation of metabolically engineered E. coli strain. Among E. coli strains WL3110, XL1-Blue, and BL21(DE3), recombinant E. coli XL1-Blue strain expressing PhaC1437 and Pct540 produced P(76.4mol%2HB-co-23.6mol%LA) to the highest content of 88 wt% when it was cultured in a chemically defined medium containing 20 g/L of glucose and 2 g/L of sodium 2-hydroxybutyrate. When recombinant E. coli XL1-Blue strain expressing PhaC1437 and Pct540 was cultured in a chemically defined medium containing 20 g/L of glucose and varying concentration of sodium 2-hydroxybutyrate, 2HB monomer fraction in P(2HB-co-LA) increased proportional to the concentration of sodium 2-hydroxybutyrate added to the culture medium. P(2HB-co-LA)] could also be produced from glucose as a sole carbon source without sodium 2-hydroxybutyrate into the culture medium. Recombinant E. coli XL1-Blue strain expressing the phaC1437, pct540, cimA3.7, and leuBCD genes together with the L. lactis Il1403 panE gene, successfully produced P(23.5mol%2HB-co-76.5mol%LA)] to the polymer content of 19.4 wt% when it cultured in a chemically defined medium containing 20 g/L of glucose. The metabolic engineering strategy reported here should be useful for the production of novel copolymer P(2HB-co-LA)]. |
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