Alcoholytic Cleavage of Polyhydroxyalkanoate Chains by Class IV Synthases Induced by Endogenous and Exogenous Ethanol

Appl Environ Microbiol. 2014 Feb;80(4):1421-9. doi: 10.1128/AEM.03576-13. Epub 2013 Dec 13.

Abstract

Polyhydroxyalkanoate (PHA)-producing Bacillus strains express class IV PHA synthase, which is composed of the subunits PhaR and PhaC. Recombinant Escherichia coli expressing PHA synthase from Bacillus cereus strain YB-4 (PhaRCYB-4) showed an unusual reduction of the molecular weight of PHA produced during the stationary phase of growth. Nuclear magnetic resonance analysis of the low-molecular-weight PHA revealed that its carboxy end structure was capped by ethanol, suggesting that the molecular weight reduction was the result of alcoholytic cleavage of PHA chains by PhaRCYB-4 induced by endogenous ethanol. This scission reaction was also induced by exogenous ethanol in both in vivo and in vitro assays. In addition, PhaRCYB-4 was observed to have alcoholysis activity for PHA chains synthesized by other synthases. The PHA synthase from Bacillus megaterium (PhaRCBm) from another subgroup of class IV synthases was also assayed and was shown to have weak alcoholysis activity for PHA chains. These results suggest that class IV synthases may commonly share alcoholysis activity as an inherent feature.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Acyltransferases / genetics
  • Acyltransferases / metabolism*
  • Bacillus cereus / enzymology*
  • Bacillus cereus / genetics
  • Bacillus megaterium / enzymology
  • Escherichia coli / genetics
  • Escherichia coli / metabolism*
  • Ethanol / metabolism*
  • Hydrolysis
  • Magnetic Resonance Spectroscopy
  • Polyhydroxyalkanoates / metabolism*
  • Recombinant Proteins / genetics
  • Recombinant Proteins / metabolism

Substances

  • Polyhydroxyalkanoates
  • Recombinant Proteins
  • Ethanol
  • Acyltransferases
  • poly(3-hydroxyalkanoic acid) synthase