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Related Experiment Videos

Polymeric Beta-Hydroxyalkanoates from Environmental Samples and Bacillus megaterium.

R H Findlay1, D C White

  • 1Department of Biological Science, Florida State University, Tallahassee, Florida 32306.

Applied and Environmental Microbiology
|January 1, 1983
PubMed
Summary

Poly-beta-hydroxyalkanoate (PHA) is a complex polymer found in marine sediments and bacteria. Its composition can indicate the metabolic status of microbial communities.

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Area of Science:

  • Microbiology
  • Environmental Science
  • Analytical Chemistry

Background:

  • Poly-beta-hydroxybutyrate, a prokaryotic storage polymer, is a mixed polymer of short-chain beta-hydroxy fatty acids.
  • This polymer is known as poly-beta-hydroxyalkanoate (PHA).

Purpose of the Study:

  • To develop a method for quantitative recovery and analysis of PHA.
  • To analyze the composition of PHA in marine sediments and bacterial monocultures.
  • To investigate the relationship between PHA concentration and microbial activity.

Main Methods:

  • Quantitative recovery of PHA using a modified Soxhlet extractor.
  • Purification of PHA with ethanol and diethyl ether.
  • Analysis of PHA composition by capillary gas-liquid chromatography after acid-catalyzed ethanolysis.

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  • Detection limit of 100 fmol achieved with excellent reproducibility.

    • Main Results:

    • PHA from marine sediments contained at least 11 different short-chain beta-hydroxy acids.
    • PHA from Bacillus megaterium monocultures was a complex mixture of beta-hydroxy acids (C4-C8).
    • Mechanical disturbance and chelators increased PHA components relative to bacterial biomass.
    • Clymenella sp. (annelid worm) induced decreases in PHA and altered its component proportions.

    Conclusions:

    • PHA is a complex mixture of beta-hydroxy acids, not just poly-beta-hydroxybutyrate.
    • The developed method allows sensitive and reproducible PHA analysis.
    • PHA concentration relative to bacterial biomass reflects the recent metabolic status of the microbiota.