Journal of Microbiology and Biotechnology

The Korean Society for Microbiology and Biotechnology (한국미생물·생명공학회)
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Analysis of Poly(3-Hydroxybutyrate) Granule-Associated Proteome in Recombinant Escherichia coli

  • Han Mee-Jung (Metabolic and Biomolecular Engineering National Research Laboratory, Department of Chemical & Biomolecular Engineering, BioProcess Engineering Research Center, and Bioinformatics Research Center, Korea Advanced Institute of Science and Technology) ;
  • Park Si-Jae (Metabolic and Biomolecular Engineering National Research Laboratory, Department of Chemical & Biomolecular Engineering, BioProcess Engineering Research Center, and Bioinformatics Research Center, Korea Advanced Institute of Science and Technology) ;
  • Lee Jeong-Wook (Metabolic and Biomolecular Engineering National Research Laboratory, Department of Chemical & Biomolecular Engineering, BioProcess Engineering Research Center, and Bioinformatics Research Center, Korea Advanced Institute of Science and Technology) ;
  • Min Byoung-Hoon (Cell Biology Laboratory, Department of Biology, Hallym University) ;
  • Lee Sang-Yup (Metabolic and Biomolecular Engineering National Research Laboratory, Department of Chemical & Biomolecular Engineering, BioProcess Engineering Research Center, and Bioinformatics Research Center, Korea Advanced Institute of Science and Technology) ;
  • Kim Soo-Jin (Metabolic and Biomolecular Engineering National Research Laboratory, Department of Chemical & Biomolecular Engineering, BioProcess Engineering Research Center, and Bioinformatics Research Center, Korea Advanced Institute of Science and Technology) ;
  • Yoo Jong-Shin (Korea Basic Science Institute)
  • Published : 2006.06.01
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Abstract

Poly(3-hydroxybutyrate) [P(3HB)] is a microbial polyester intracellularly accumulated as distinct granules in numerous microorganisms as an energy and carbon storage material. Recombinant Escherichia coli harboring the heterologous P(3HB) biosynthesis genes accumulates large amounts of P(3HB) granules, yet the granule-associated proteins have not been identified. Therefore, this study reports on an analysis of the P(3HB) granule-associated proteome in recombinant E. coli. Fiye proteins out of 7 spots identified were found to be involved in functions of translation, heat-stress responses, and P(3HB) biosynthesis. Two of the major granule-associated proteins, IbpA/B, which are already known to bind to recombinant proteins forming inclusion bodies in E. coli, were further analyzed. Immunoblotting and immunoelectron microscopic studies with IbpA/B antibodies clearly demonstrated the binding and localization of IbpA/B to P(3HB) granules. IbpA/B seemed to play an important role in recombinant E. coli producing P(3HB) by stabilizing the interface between the hydrophobic P(3HB) granules and the hydrophilic cytoplasm. Thus, IbpA/B were found to act like phasins in recombinant E. coli, as they are the major proteins bound to the P(3HB) granules, affect the morphology of the granules, and reduce the amount of cytosolic proteins bound to the P(3HB) granules.

Keywords

References

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