Metagenomics pp 237-248 | Cite as

Methods for the Isolation of Genes Encoding Novel PHA Metabolism Enzymes from Complex Microbial Communities

  • Jiujun Cheng
  • Ricardo Nordeste
  • Maria A. Trainer
  • Trevor C. CharlesEmail author
Part of the Methods in Molecular Biology book series (MIMB, volume 1539)


Development of different PHAs as alternatives to petrochemically derived plastics can be facilitated by mining metagenomic libraries for diverse PHA cycle genes that might be useful for synthesis of bio-plastics. The specific phenotypes associated with mutations of the PHA synthesis pathway genes in Sinorhizobium meliloti and Pseudomonas putida, allows the use of powerful selection and screening tools to identify complementing novel PHA synthesis genes. Identification of novel genes through their function rather than sequence facilitates the functional proteins that may otherwise have been excluded through sequence-only screening methodology. We present here methods that we have developed for the isolation of clones expressing novel PHA metabolism genes from metagenomic libraries.

Key words

PHA/PHB pathway Sinorhizobium meliloti Pseudomonas putida Microbial community gene libraries Phenotypic complementation 


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Copyright information

© Springer Science+Business Media LLC 2017

Authors and Affiliations

  • Jiujun Cheng
    • 1
  • Ricardo Nordeste
    • 1
  • Maria A. Trainer
    • 1
  • Trevor C. Charles
    • 1
    Email author
  1. 1.Department of BiologyUniversity of WaterlooWaterlooCanada

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