Precise control of repeating unit composition in biodegradable poly(3-hydroxyalkanoate) polymers synthesized by Escherichia coli(MICROBIAL PHYSIOLOGY AND BIOTECHNOLOGY)

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概要

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• The composition of medium-chain-length (MCL) poly(3-hydroxyalkanoate) (PHA) biopolymers is normally an uncontrollable random mixture of repeating units with differing side chain lengths. Attempts to generate MCL PHA homopolymers and control repeating unit composition have been published in native PHA-produdng organisms but have limited ranges for the different sizes of repeating units that can be synthesized. In this study, a new Escherichia coil-based system that exhibits control over repeating unit composition for both MCL PHAs and short-chain-length (SCL) PHAs has been developed, covering an unprecedented range of repeating units. The fadB and fadJ genes from the β-oxidation pathway were eliminated from the chromosome of E. coil LS5218. The subsequent blockage in β-oxidation caused a buildup of enoyl-CoA intermediates, which were converted to PHAs by an (R)-specffic enoyl-CoA hydratase (PhaJ4) and PHA synthase [PhaC1(STQK)J expressed froma plasmid DNA construct. Fatty acid substrates were converted to PHAs with repeating units equal in the number of carbon atoms to the fatty acid substrate. The broad substrate specificities of the PhaJ4 and PhaC1(STQK) enzymes allowed for the production of homopolymers with strict control over the repeating unit composition from substrates of four to twelve carbons in length. Polymers were purified and analyzed by GC,GC-MS. and NMR for structural composition and by DSC, TGA, and GPC for thermal and physical characteristics. This study marks the development of the first single biological system to achieve consistent repeating unit control over such a broad range of repeating units in PHAs.

著者

• Wang Qin

  State Key Laboratory Of Chemical Resource Engineering

• Nomura Christopher

  State University Of New York College Of Environmental Science And Forestry

• Wang Qin

  State University Of New York College Of Environmental Science And Forestry

• Tappel Ryan

  State University Of New York College Of Environmental Science And Forestry 1 Forestry Drive Chemistry Department

• Tappel Ryan

  State University of New York College of Environmental Science and Forestry

• Wang Qin

  State Key Laboratory of Cellular Stress Biology and Key Laboratory of the Ministry of Education for Coastal and Wetland Ecosystems, School of Life Sciences, Xiamen University

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