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Related Experiment Videos

Universal immunoprobe for (per)chlorate-reducing bacteria.

Susan M O'Connor1, John D Coates

  • 1Department of Microbiology, Southern Illinois University, Carbondale, Illinois 62901, USA.

Applied and Environmental Microbiology
|June 1, 2002
PubMed
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Researchers developed a sensitive immunoprobe to detect diverse anaerobic chlorate-reducing bacteria (ClRB). This probe targets the conserved chlorite dismutase (CD) enzyme, enabling environmental assessment of these crucial microorganisms.

Area of Science:

  • Environmental microbiology
  • Biochemistry
  • Molecular biology

Background:

  • Anaerobic microorganisms reduce chlorate or perchlorate [(per)chlorate] for growth.
  • Dechloromonas and Dechlorosoma groups are dominant (per)chlorate-reducing bacteria (ClRB).
  • Chlorite dismutation, mediated by chlorite dismutase (CD), is a conserved step in (per)chlorate reduction.

Purpose of the Study:

  • To develop a specific and sensitive probe for identifying ClRB in environmental samples.
  • To leverage the conserved nature of the chlorite dismutase (CD) enzyme for this purpose.

Main Methods:

  • Polyclonal antibodies were raised against purified CD from Dechloromonas agitata.
  • Antiserum activity was assessed using dot blot analysis with purified CD, cell lysates, and whole cells.

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  • The probe's specificity was tested against diverse ClRB and related non-ClRB organisms.

    • Main Results:

    • The developed antiserum showed high affinity and sensitivity for CD, active at high dilutions.
    • The immunoprobe detected ClRB grown anaerobically with (per)chlorate, but not aerobically grown cells.
    • The probe successfully identified diverse ClRB from Proteobacteria subclasses, irrespective of phylogenetic affiliation.
    • No cross-reactivity was observed with closely related organisms unable to reduce (per)chlorate.

    Conclusions:

    • A highly specific and sensitive immunoprobe for ClRB has been developed, targeting the conserved CD enzyme.
    • This tool allows for the assessment of dissimilatory (per)chlorate-reducing populations in environmental samples.
    • The probe's effectiveness across diverse ClRB phylogenetic groups highlights the utility of targeting conserved enzymes.