title: Formation of short chain length/medium chain length polyhydroxyalkanoate copolymers by fatty acid beta-oxidation inhibited Ralstonia eutropha.
abstract: Ralstonia eutropha has been considered as a bacterium, incorporating hydroxyalkanoates of less than six carbons only into polyhydroxyalkanoates (PHAs). Cells of the wild type cultivated with sodium octanoate as the carbon source in the presence of the fatty acid beta-oxidation inhibitor sodium acrylate synthesized PHAs composed of the medium chain length hydroxyalkanoates (3HA(MCL)) 3-hydroxyhexanoate (3HHx) and 3-hydroxyoctanoate (3HO) as well as of 3-hydroxybutyrate and 3-hydroxyproprionate as revealed by gas chromatography, (1)H NMR spectroscopy, and mass spectroscopy. The characterization of the polymer as a tetrapolymer was confirmed by differential solvent extraction and measurement of melting and glass transition temperature depression in the purified polymer compared to PHB. These data suggested that the R. eutropha PHA synthase is capable of incorporating longer chain substrates than suggested by previous in vitro studies. Furthermore, expression of the class II PHA synthase gene phaC1 from P. aeruginosa in R. eutropha resulted in the accumulation of PHAs consisting of 3HA(MCL) contributing about 3-5% to cellular dry weight. These PHAs were composed of nearly equal molar fractions of 3HO and 3-hydroxydecanoate (3HD) with traces of 3HHx. These data indicated that 3HA(MCL)-CoA thioesters were diverted from the fatty acid beta-oxidation pathway towards PHA biosynthesis in recombinant R. eutropha.
authors.affiliation: The Procter and Gamble Company, Cincinnati, Ohio 45253-8707, USA.
authors.forename: Phillip R
authors.initials: PR
authors.lastname: Green
authors.forename: Joe
authors.initials: J
authors.lastname: Kemper
authors.forename: Lee
authors.initials: L
authors.lastname: Schechtman
authors.forename: Ling
authors.initials: L
authors.lastname: Guo
authors.forename: Mike
authors.initials: M
authors.lastname: Satkowski
authors.forename: Silke
authors.initials: S
authors.lastname: Fiedler
authors.forename: Alexander
authors.initials: A
authors.lastname: Steinbüchel
authors.forename: Bernd H A
authors.initials: BH
authors.lastname: Rehm
mesh_terms: D020155:3-Hydroxybutyric Acid / Q000378:metabolism; D000179:Acrylates / Q000494:pharmacology; D000217:Acyltransferases / Q000378:metabolism; D002478:Cells, Cultured; D002849:Chromatography, Gas; D020118:Cupriavidus necator / Q000201:enzymology* / Q000235:genetics; D004926:Escherichia coli / Q000737:chemistry / Q000201:enzymology / Q000235:genetics; D005227:Fatty Acids / Q000037:antagonists & inhibitors / Q000378:metabolism*; D008401:Gas Chromatography-Mass Spectrometry; D010084:Oxidation-Reduction; D010957:Plasmids / Q000235:genetics; D011091:Polyesters / Q000032:analysis / Q000378:metabolism*; D011108:Polymers / Q000737:chemistry / Q000378:metabolism*; D011994:Recombinant Proteins / Q000378:metabolism
chemical_list: D000179:Acrylates; D005227:Fatty Acids; D011091:Polyesters; D011108:Polymers; D011994:Recombinant Proteins; D000217:Acyltransferases; C077225:poly(3-hydroxyalkanoic acid) synthase; D020155:3-Hydroxybutyric Acid