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

  • Metabolic Engineering and Synthetic Biology
  • Published:
Journal of Industrial Microbiology & Biotechnology

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.

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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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Authors and Affiliations

  1. Chemical Engineering, Queen’s University, Kingston, ON, K7L 3N6, Canada

    Minh Tri Vo & Bruce Ramsay

  2. Biology, Queen’s University, 116 Barrie St., Kingston, ON, K7L 3N6, Canada

    Kenton Ko

  3. Polyferm Canada, 4530 Camden-Portland Bdry Rd, RR#1, Harrowsmith, ON, K0H 1V0, Canada

    Bruce Ramsay

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  1. Minh Tri Vo
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Correspondence to Bruce Ramsay.

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Vo, M.T., Ko, K. & Ramsay, B. Carbon-limited fed-batch production of medium-chain-length polyhydroxyalkanoates by a phaZ-knockout strain of Pseudomonas putida KT2440. J Ind Microbiol Biotechnol 42, 637–646 (2015). https://doi.org/10.1007/s10295-014-1574-5

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  • DOI: https://doi.org/10.1007/s10295-014-1574-5

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