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10:21
Expression, Purification, Crystallization, and Enzyme Assays of Fumarylacetoacetate Hydrolase Domain-Containing Proteins
Published on: June 20, 2019
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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
Summary
Researchers discovered novel thermostable hydrolases from extreme environments using advanced screening methods. These enzymes offer new possibilities for industrial processes and environmental applications.
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.
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