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Related Experiment Video

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Expression, Purification, Crystallization, and Enzyme Assays of Fumarylacetoacetate Hydrolase Domain-Containing Proteins
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Discovering novel hydrolases from hot environments.

Roland Wohlgemuth1, Jennifer Littlechild2, Daniela Monti3

  • 1Sigma-Aldrich, Member of Merck Group, Industriestrasse 25, CH-9470 Buchs, Switzerland; Institute of Technical Biochemistry, Lodz University of Technology, ul. Stefanowskiego 4/10, 90-924 Lodz, Poland.

Biotechnology Advances
|September 30, 2018
PubMed
Summary

Researchers discovered novel thermostable hydrolases from extreme environments using advanced screening methods. These enzymes offer new possibilities for industrial processes and environmental applications.

Keywords:
BiocatalysisEnrichmentEnzyme characterizationEnzyme screeningGene expressionGenomicsHydrolasesMetagenomicsSequencingThermophiles

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Area of Science:

  • Enzyme discovery and engineering
  • Biotechnology
  • Environmental microbiology

Background:

  • Novel hydrolases from extreme environments are valuable for industrial applications.
  • Existing enzymes and processes can be improved by thermostable hydrolases.
  • Systematic screening methods are needed for discovering enzymes with desired functions.

Purpose of the Study:

  • To develop and integrate technologies for discovering, identifying, and characterizing novel thermostable hydrolases.
  • To mine hot terrestrial and marine environments for new enzymes.
  • To establish new tools for enzyme discovery and functional screening.

Main Methods:

  • Mining hot springs for environmental samples and enrichment cultures.
  • High-throughput sequencing (metagenomes, genomes, transcriptomes).
  • Bioinformatics analysis using the Anastasia platform and whole-cell bioreporter-based functional screening.

Main Results:

  • Hundreds of samples and thousands of cultures were generated.
  • 15 hot spring metagenomes and several isolate genomes/transcriptomes were sequenced.
  • Approximately 100 potential novel hydrolases were identified, with over a dozen characterized (e.g., carboxylesterases, cellulases).

Conclusions:

  • The HotZyme project successfully developed a workflow for discovering and characterizing novel thermostable hydrolases.
  • The identified enzymes possess valuable biochemical and structural properties for various applications.
  • The generated data and samples provide a foundation for future enzyme discovery.