title: Characterization of polyhydroxyalkanoate (PHA) synthase derived from Delftia acidovorans DS-17 and the influence of PHA production in Escherichia coli.
abstract: 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.
authors: Department of Innovative and Engineered Materials, Tokyo Institute of Technology, 4259 Nagatsuta, Midori-ku, Yokohama 226-8502, Japan.
authors: Ayaka
authors: A
authors: Hiroe
authors: Kazunori
authors: K
authors: Ushimaru
authors: Takeharu
authors: T
authors: Tsuge
mesh_terms: D000217:Acyltransferases / Q000235:genetics / Q000378:metabolism*; D020118:Cupriavidus necator / Q000201:enzymology; D020605:Delftia acidovorans / Q000201:enzymology*; D003902:Detergents; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D006885:Hydroxybutyrates / Q000378:metabolism*; D017830:Octoxynol / Q000737:chemistry; D011091:Polyesters / Q000378:metabolism*; D054813:Polyhydroxyalkanoates / Q000096:biosynthesis; D000094122:Polyhydroxybutyrates
chemical_list: D000217:Acyltransferases; D003902:Detergents; D006885:Hydroxybutyrates; D017830:Octoxynol; D011091:Polyesters; D054813:Polyhydroxyalkanoates; C003182:poly-beta-hydroxybutyrate; D000094122:Polyhydroxybutyrates; C077225:poly(3-hydroxyalkanoic acid) synthase