Novel polyhydroxyalkanoate copolymers produced in Pseudomonas putida by metagenomic polyhydroxyalkanoate synthases

Appl Microbiol Biotechnol. 2016 Sep;100(17):7611-27. doi: 10.1007/s00253-016-7666-6. Epub 2016 Jun 23.

Abstract

Bacterially produced biodegradable polyhydroxyalkanoates (PHAs) with versatile properties can be achieved using different PHA synthases (PhaCs). This work aims to expand the diversity of known PhaCs via functional metagenomics and demonstrates the use of these novel enzymes in PHA production. Complementation of a PHA synthesis-deficient Pseudomonas putida strain with a soil metagenomic cosmid library retrieved 27 clones expressing either class I, class II, or unclassified PHA synthases, and many did not have close sequence matches to known PhaCs. The composition of PHA produced by these clones was dependent on both the supplied growth substrates and the nature of the PHA synthase, with various combinations of short-chain-length (SCL) and medium-chain-length (MCL) PHA. These data demonstrate the ability to isolate diverse genes for PHA synthesis by functional metagenomics and their use for the production of a variety of PHA polymer and copolymer mixtures.

Keywords: Bioplastics; Functional metagenomics; Polyhydroxyalkanoate; Polyhydroxyalkanoate synthase; Pseudomonas putida; Sinorhizobium meliloti; Soil metagenomic library.

MeSH terms

  • Acyltransferases / genetics*
  • Acyltransferases / metabolism*
  • Base Sequence
  • Cloning, Molecular
  • DNA, Bacterial / genetics
  • Gene Expression Regulation, Bacterial
  • Gene Library
  • Metagenomics
  • Polyhydroxyalkanoates / metabolism*
  • Pseudomonas putida / enzymology*
  • Pseudomonas putida / genetics*
  • Pseudomonas putida / metabolism
  • Sequence Analysis, DNA

Substances

  • DNA, Bacterial
  • Polyhydroxyalkanoates
  • Acyltransferases
  • poly(3-hydroxyalkanoic acid) synthase