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Novel polyhydroxyalkanoate copolymers produced in Pseudomonas putida by metagenomic polyhydroxyalkanoate synthases

  • Applied genetics and molecular biotechnology
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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.

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Acknowledgments

We are very grateful to Ricardo Nordeste for assisting with GC-MS analysis of PHA. We wish to thank Dr. Bruce Ramsay (Polyferm Canada) for providing PHA standards and Dr. Josh Neufeld for use of the FilterMax F5 Multi-mode microplate reader.

Authors’s contributions

JC and TCC conceived and designed this study, carried out all the experiments, and prepared the manuscript. Both authors read and approved the final manuscript.

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Authors and Affiliations

  1. Department of Biology and Centre for Bioengineering and Biotechnology, University of Waterloo, Waterloo, ON, N2L 3G1, Canada

    Jiujun Cheng & Trevor C. Charles

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  1. Jiujun Cheng
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  2. Trevor C. Charles
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Correspondence to Trevor C. Charles.

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Funding

This work was financially supported by a New Directions grant from Ontario Ministry of Agriculture, Food, and Rural Affairs (award number 381646-09), by a Strategic Projects grant from the Natural Sciences and Engineering Research Council of Canada (award number ND2012-1679), and by Genome Canada, through the Applied Genomics Research in Bioproducts or Crops (ABC) program for the grant titled “Microbial Genomics for Biofuels and Co-Products from Biorefining Processes” awarded to David Levin and Richard Sparling.

Conflict of interest

Trevor Charles received partial financial support for presenting some of this material at the Society for Industrial Microbiology and Biotechnology Annual Meeting in 2014 and 2015. Jiujun Cheng declares no conflict of interest.

Ethical statement

The authors certify that this manuscript has not been published previously, and not under consideration for publication elsewhere, in whole or in part. No data have been fabricated or manipulated (including images), and no data, text, or theories by others are presented as if they were the authors’ own. Consent to submit has been received explicitly from all the authors listed. And authors whose names appear on the submission have contributed sufficiently to the scientific work and therefore share collective responsibility and accountability for the results. This article does not contain any studies with human participants or animals performed by any of the authors.

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Cheng, J., Charles, T.C. Novel polyhydroxyalkanoate copolymers produced in Pseudomonas putida by metagenomic polyhydroxyalkanoate synthases. Appl Microbiol Biotechnol 100, 7611–7627 (2016). https://doi.org/10.1007/s00253-016-7666-6

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