Characterization of polyhydroxyalkanoate synthases from the marine bacterium Neptunomonas concharum JCM17730

Abstract

Neptunomonas concharum JCM17730 was isolated from an ark clam sample and characterized as a mesophilic bacterium. The genome of N. concharum JCM17730 contains thirteen genes related to polyhydroxyalkanoates (PHA) metabolism. Three PHA synthase encoding genes were identified, and phylogenetic analysis of enzyme sequences suggested the presence of two class I PHA synthases and one class III PHA synthase. The PHA synthases of N. concharum were heterologously expressed with acetyl-CoA acetyltransferase and acetoacetyl-CoA reductase in Escherichia coli to confirm the catalytic activity of each PHA synthase. Recombinants harboring different PHA synthase exhibit important distinctions in poly-3-hydroxybutyrate synthesis ability under various temperatures. Decreased cultivation temperature (≤30 °C) significantly improved PHB titer and content. This is the first report on characterization of PHA synthases from the marine genus Neptunomonas and would provide molecular basis for PHA production using Neptunomonas species.