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Distribution and Selection of Poly-3-Hydroxybutyrate Production Capacity in Methanotrophic Proteobacteria

Abstract

Methanotrophs are known to produce poly-3-hydroxybutyrate (PHB), but there is conflicting evidence in the literature as to which genera produce the polymer. We screened type I and II proteobacterial methanotrophs that use the ribulose monophosphate and serine pathways for carbon assimilation, respectively, for both phaC, which encodes for PHB synthase, and the ability to produce PHB under nitrogen-limited conditions. Twelve strains from six different genera were evaluated. All type I strains tested negative for phaC and PHB production; all Type II strains tested positive for phaC and PHB production. In order to identify conditions that favor PHB production, we also evaluated a range of selection conditions using a diverse activated sludge inoculum. Use of medium typically recommended for methanotroph enrichment led to enrichments dominated by type I methanotrophs. Conditions that were selected for enrichments dominated by PHB-producing Type II methanotrophs were: (1) use of nitrogen gas as the sole nitrogen source in the absence of copper, (2) use of a dilute mineral salts media in the absence of copper, and (3) use of media prepared at pH values of 4–5.

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Acknowledgments

We thank Dr. Lisa Stein and the Carbon-one Oxidation Network for the phaC gene sequence for M. trichosporium OB3b. We also thank Holly Sewell for her help in the laboratory.

This work was supported by a Graduate Research Fellowship from the National Science Foundation to AJP and by the California Environmental Protection Agency, Department of Toxic Substances Control under contract 07T3451.

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Correspondence to Craig S. Criddle.

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Pieja, A.J., Rostkowski, K.H. & Criddle, C.S. Distribution and Selection of Poly-3-Hydroxybutyrate Production Capacity in Methanotrophic Proteobacteria. Microb Ecol 62, 564–573 (2011) doi:10.1007/s00248-011-9873-0

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Keywords

  • Terminal Restriction Fragment Length Polymorphism
  • Methanotrophs
  • Sole Nitrogen Source
  • pmoA Gene
  • Methylocystis