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Related Experiment Videos

Intracellular depolymerase functionality and location in Pseudomonas oleovorans inclusions containing

E S Stuart1, L J Foster, R W Lenz

  • 1Dept. of Biochemistry and Molecular Biology, University of Massachusetts, Lederle Graduate Research Center, Amherst 01003-4505, USA.

International Journal of Biological Macromolecules
|October 1, 1996
PubMed
Summary
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Pseudomonas oleovorans stores polyester in inclusion bodies. A 32 kDa depolymerase protein regulates polyester breakdown, increasing when carbon is scarce to ensure cell survival.

Area of Science:

  • Microbiology
  • Biochemistry
  • Polymer Science

Background:

  • Microbial poly-3-hydroxyoctanoate (PHO) inclusion bodies are complex structures.
  • The roles of components in PHO production and utilization are not fully understood.

Purpose of the Study:

  • Identify the depolymerase protein in PHO inclusion bodies.
  • Investigate the metabolic regulation of depolymerase and polymerase during polyester utilization.

Main Methods:

  • Electron microscopy
  • SDS-PAGE
  • Gel filtration chromatography
  • In vitro depolymerase activity assays

Main Results:

  • A ~32 kDa protein was identified as the depolymerase.

Related Experiment Videos

  • Depolymerase concentration increased, while polymerase concentration decreased upon carbon starvation.
  • Depolymerase association with granules is metabolically regulated.

    • Conclusions:

    • Substrate concentration manipulation can alter inclusion-associated proteins and polyester accumulation.
    • Metabolic regulation controls polyester mobilization via depolymerase activity.