title: Novel polyhydroxyalkanoate copolymers produced in Pseudomonas putida by metagenomic polyhydroxyalkanoate synthases.
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.
authors.affiliation: Department of Biology and Centre for Bioengineering and Biotechnology, University of Waterloo, Waterloo, ON, N2L 3G1, Canada.
authors.forename: Jiujun
authors.initials: J
authors.lastname: Cheng
authors.affiliation: Department of Biology and Centre for Bioengineering and Biotechnology, University of Waterloo, Waterloo, ON, N2L 3G1, Canada. tcharles@uwaterloo.ca.
authors.forename: Trevor C
authors.initials: TC
authors.lastname: Charles
mesh_terms: D000217:Acyltransferases / Q000235:genetics* / Q000378:metabolism*; D001483:Base Sequence; D003001:Cloning, Molecular; D004269:DNA, Bacterial / Q000235:genetics; D015964:Gene Expression Regulation, Bacterial; D015723:Gene Library; D056186:Metagenomics; D054813:Polyhydroxyalkanoates / Q000378:metabolism*; D016958:Pseudomonas putida / Q000201:enzymology* / Q000235:genetics* / Q000378:metabolism; D017422:Sequence Analysis, DNA
chemical_list: D004269:DNA, Bacterial; D054813:Polyhydroxyalkanoates; D000217:Acyltransferases; C077225:poly(3-hydroxyalkanoic acid) synthase
keywords: Bioplastics; Functional metagenomics; Polyhydroxyalkanoate; Polyhydroxyalkanoate synthase; Pseudomonas putida; Sinorhizobium meliloti; Soil metagenomic library