Functional metagenomics using Pseudomonas putida expands the known diversity of polyhydroxyalkanoate synthases and enables the production of novel polyhydroxyalkanoate copolymers

Functional metagenomics using Pseudomonas putida expands the known diversity of 1 polyhydroxyalkanoate synthases and enables the production of novel polyhydroxyalkanoate 2 copolymers 3 4 14 15. CC-BY 4.0 International license peer-reviewed) is the author/funder. It is made available under a The copyright holder for this preprint (which was not. Abstract 17 Bacterially produced biodegradable polyhydroxyalkanoates with versatile properties can be achieved 18 using different PHA synthase enzymes. This work aims to expand the diversity of known PHA 19 synthases via functional metagenomics, and demonstrates the use of these novel enzymes in PHA 20 production. Complementation of a PHA synthesis deficient Pseudomonas putida strain with a soil 21 metagenomic cosmid library retrieved 27 clones expressing either Class I, Class II or unclassified 22 PHA synthases, and many did not have close sequence matches to known PHA synthases. The 23 composition of PHA produced by these clones was dependent on both the supplied growth substrates 24 and the nature of the PHA synthase, with various combinations of SCL-and MCL-PHA. These data 25 demonstrate the ability to isolate diverse genes for PHA synthesis by functional metagenomics, and 26 their use for the production of a variety of PHA polymer and copolymer mixtures.