Characterization of polyhydroxyalkanoate (PHA) synthase derived from Delftia acidovorans DS-17 and the influence of PHA production in Escherichia coli(ENZYMOLOGY, PROTEIN ENGINEERING, AND ENZYME TECHNOLOGY)
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概要
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Heterologous expression of polyhydroxyalkanoate (PHA) synthase from Delftia acidovorans DS-17 (PhaC_<Da>) in Escherichia coli JM109 leads to effective production of high-molecular-weight poly[(R)-3-hydroxybutyrate] [P(3HB)]. This study examined the effect of PhaC_<Da> expression on P(3HB) production in E. coli JM109 (Da strain) by comparing with the strain expressing PHA synthase (PhaC_<Re>) from Ralstonia eutropha (Re strain). First, the kinetic properties of PhaC_<Da> were investigated. Among the five detergents examined, Triton X-100 remarkably activated PhaC_<Da>, as well as PhaC_<Re>. The affinity of PhaC_<Da> for its substrate was lower than that of PhaC_<Re>, whereas the maximum reaction rate of PhaC_<Da> was higher than that of PhaC_<Re>. However, the kinetic differences were not likely to influence P(3HB) production in the cells. Under conditions of P(3HB) production, the translational levels of monomer-supplying enzymes (PhaA and PhaB) were similar in both the Da and Re strains, whereas PhaC exhibited different expression levels: the abundance of soluble PhaC_<Da> was lower than that of soluble PhaC_<Re>. This observation suggests that the production of high-molecular-weight P(3HB) by the Da strain would be attributed to the low amounts of active PhaC_<Da> in the cells.
- 2013-06-00
著者
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TSUGE TAKEHARU
Department of Food Science and Technology, Faculty of Agriculture, Kyushu University
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USHIMARU Kazunori
Department of Innovative and Engineered Materials, Tokyo Institute of Technology
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Hiroe Ayaka
Department of Innovative and Engineered Materials, Tokyo Institute of Technology
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