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A method for screening polyphosphate-accumulating mutants which remove phosphate efficiently from synthetic

Tomohiro Morohoshi1, Tomohiro Yamashita, Junichi Kato

  • 1Department of Molecular Biotechnology, Graduate School of Advanced Sciences of Matter, Hiroshima University, Higashi-Hiroshima, Hiroshima 739-8530, Japan.

Journal of Bioscience and Bioengineering
|October 20, 2005
PubMed
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Researchers identified a simple screening method for bacteria that accumulate polyphosphate (polyp), a key component in biological phosphorus removal from wastewater. This technique utilizes a specific chemical indicator on agar plates for easy mutant identification.

Area of Science:

  • Environmental Microbiology
  • Biotechnology
  • Wastewater Treatment

Background:

  • Biological phosphorus removal in wastewater relies on bacteria accumulating inorganic polyphosphate (polyp).
  • A phoU mutation in Escherichia coli was previously shown to induce polyp accumulation.
  • A screening method using 5-bromo-4-chloro-3-indolyl-phosphate (X-P(i)) and N-methyl-N'-nitro-N-nitrosoguanidine (NTG) mutagenesis was developed for E. coli.

Purpose of the Study:

  • To evaluate the utility of the established screening method for identifying polyp-accumulating bacterial mutants from environmental samples.
  • To broaden the application of a cost-effective screening technique for enhanced biological phosphorus removal.

Main Methods:

  • Bacterial strains were isolated from soil and activated sludge samples.

Related Experiment Videos

  • Mutagenesis was induced using N-methyl-N'-nitro-N-nitrosoguanidine (NTG).
  • Mutants were screened on agar plates containing 5-bromo-4-chloro-3-indolyl-phosphate (X-P(i)) to identify polyphosphate accumulators.

    • Main Results:

    • The screening method successfully identified polyphosphate-accumulating mutants from diverse bacterial strains.
    • The method proved effective for bacterial strains isolated from both soil and activated sludge.
    • The ease of screening facilitates the discovery of new bacterial candidates for wastewater treatment.

    Conclusions:

    • The described screening method is a versatile and effective tool for identifying polyphosphate-accumulating bacteria from environmental sources.
    • This technique can accelerate the discovery and development of microbial solutions for biological phosphorus removal in wastewater treatment.
    • The findings support the use of this method for isolating novel bacteria with enhanced phosphorus removal capabilities.