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Bacillus caldolyticus prs gene encoding phosphoribosyl-diphosphate synthase

B N Krath1, B Hove-Jensen

  • 1Center for Enzyme Research, University of Copenhagen, Denmark.

Gene
|October 17, 1996
PubMed
Summary

Bacillus caldolyticus PRPP synthase, encoded by the prs gene, shows high similarity across species and is heat-resistant. This thermophile

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Area of Science:

  • Microbiology
  • Molecular Biology
  • Biochemistry

Background:

  • The prs gene encodes phosphoribosyl-diphosphate (PRPP) synthase, an essential enzyme.
  • Thermostable enzymes are valuable for industrial applications.
  • Comparative genomics reveals gene organization and evolutionary relationships.

Purpose of the Study:

  • To clone and sequence the prs gene and flanking regions from the thermophile Bacillus caldolyticus.
  • To investigate the co-transcription of the gcaD-prs-ctc gene cluster.
  • To characterize the properties of Bacillus caldolyticus PRPP synthase.

Main Methods:

  • Gene cloning and DNA sequencing.
  • Reverse transcription and polymerase chain reaction (RT-PCR) for co-transcription analysis.
  • Enzyme activity assays and heat stability tests.
  • Heterologous expression in E. coli.

Main Results:

  • The gcaD-prs-ctc gene cluster was identified and sequenced in B. caldolyticus.
  • Evidence for co-transcription of the three genes was established.
  • B. caldolyticus PRPP synthase exhibited high similarity to homologous enzymes across diverse species.
  • Recombinant B. caldolyticus PRPP synthase showed significantly higher activity and thermostability compared to B. subtilis PRPP synthase.

Conclusions:

  • The gcaD, prs, and ctc genes form a co-transcribed unit in B. caldolyticus.
  • B. caldolyticus PRPP synthase is a highly stable enzyme with potential biotechnological applications.
  • Comparative analysis highlights the evolutionary conservation and unique adaptations of PRPP synthase.

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