TY - JOUR T1 - Functional metagenomics using <em>Pseudomonas putida</em> expands the known diversity of polyhydroxyalkanoate synthases and enables the production of novel polyhydroxyalkanoate copolymers JF - bioRxiv DO - 10.1101/042705 SP - 042705 AU - Jiujun Cheng AU - Trevor C. Charles Y1 - 2016/01/01 UR - http://biorxiv.org/content/early/2016/03/07/042705.abstract N2 - Bacterially produced biodegradable polyhydroxyalkanoates with versatile properties can be achieved using different PHA synthase enzymes. This work aims to expand the diversity of known PHA synthases 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 PHA synthases. 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 SCL-and 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.HighlightsA functional metagenomic screen was developed for isolation of PHA synthesis genes using Pseudomonas putidaUse of the screen resulted in a collection of novel PHA synthesis genes that directed the production of a variety of different polyhydroxyalkanaotes, including SCL-MCL copolymersDNA sequence analysis of the cloned DNA suggests some basis for the polyhydroxyalkanoate diversity based on key amino acid residues in PHA synthase enzymesWe have demonstrated the potential for using metagenome-derived clones for production of a variety of PHAs of possible industrial utility. ER -