Carbon-limited fed-batch production of medium-chain-length polyhydroxyalkanoates by a phaZ-knockout strain of Pseudomonas putida KT2440

Metabolic Engineering and Synthetic Biology
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Abstract

A medium-chain-length poly-3-hydroxyalkanote (MCL-PHA) depolymerase knockout mutant of Pseudomonas putida KT2440 was produced by double homologous recombination. A carbon-limited shake-flask study confirmed that depolymerase activity was eliminated. Lysis of both mutant and wild-type strains occurred under these conditions. In carbon-limited, fed-batch culture, the yield of unsaturated monomers from unsaturated substrate averaged only 0.62 mol mol−1 for the phaZ minus strain compared to 0.72 mol mol−1 for the wild type. The mutant strain also produced more CO2 and less residual biomass from the same amount of carbon substrate. However, most results indicated that elimination of PHA depolymerase activity had little impact on the overall yield of biomass and PHA.

Keywords

phaZ Knockout PHA putida Depolymerase 
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Notes

Acknowledgments

The authors gratefully acknowledge the provision of plasmid pEX18Tc, pK18mobsacB and E. coli S17-1 by Professor Keith Poole at the Department of Biomedical and Molecular Sciences, Queen’s University, Kingston, Ontario, Canada, and the financial support of the Ontario Centres of Excellence.

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Copyright information

© Society for Industrial Microbiology and Biotechnology 2015

Authors and Affiliations

  1. 1.Chemical EngineeringQueen’s UniversityKingstonCanada
  2. 2.BiologyQueen’s UniversityKingstonCanada
  3. 3.Polyferm CanadaHarrowsmithCanada

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