Discovering novel hydrolases from hot environments
- Wohlgemuth, Roland [ Sigma-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 ]
- Littlechild, Jennifer [ Biocatalysis Centre, University of Exeter, The Henry Wellcome Building for Biocatalysis, Stocker Road, Exeter EX4 4QD, United Kingdom ]
- Monti, Daniela [ Istituto di Chimica del Riconoscimento Molecolare, CNR, Via Mario Bianco 9, 20131 Milano, Italy ]
- Schnorr, Kirk [ Novozymes A/S, Krogshoejvej 36, 2880 Bagsvaerd, Denmark ]
- van Rossum, Teunke [ Wageningen University and Research, Laboratory of Microbiology, Stippeneng 4, 6708 WE Wageningen, the Netherlands; Department of Bionanoscience, Kavli Institute of Nanoscience, Delft University of Technology, Van der Maasweg 9, 2629 HZ Delft, Netherlands ]
- Siebers, Bettina [ Molecular Enzyme Technology and Biochemistry, Biofilm Centre, Faculty of Chemistry, University of Duisburg-Essen, Universitaetsstr. 5, S05 V03 F41, 45141 Essen, Germany ]
- Menzel, Peter [ Department of Biology, University of Copenhagen, Ole Maaløes Vej 5, 2200 København, Denmark; Max Delbrück Center for Molecular Medicine, Robert-Rössle-Straße 10, 13125 Berlin, Germany ]
- Kublanov, Ilya V. [ Extremophiles Metabolism Laboratory, Winogradsky Institute of Microbiology, Research Center of Biotechnology, RAS, Prospekt 60 Letiya Oktyabrya 7, Moscow, Russia ]
- Rike, Anne Gunn [ Norwegian Geotechnical Institute, Sognsveien 72, N-0855 Oslo, Norway; Standard Norge, Mustads vei 1, 0283 Oslo, Norway ]
- Skretas, Georgios [ Institute of Biology, Medicinal Chemistry & Biotechnology, National Hellenic Research Foundation, 48 Vassileos Constantinou Ave, 11635 Athens, Greece ]
- Szabo, Zalan [ MicroDish BV, Padualaan 8, 3584 CH Utrecht, the Netherlands; U-Protein Express BV, Yalelaan 62, 3584 CH Utrecht, the Netherlands ]
- Peng, Xu [ Department of Biology, University of Copenhagen, Ole Maaløes Vej 5, 2200 København, Denmark ]
- Young, Mark J. [ Montana State University: Plant Sciences & Plant Pathology ]
Novel hydrolases from hot and other extreme environments showing appropriate performance and/or novel functionalities and new approaches for their systematic screening are of great interest for developing new processes, for improving safety, health and environment issues. Existing processes could benefit as well from their properties. The workflow, based on the HotZyme project, describes a multitude of technologies and their integration from discovery to application, providing new tools for discovering, identifying and characterizing more novel thermostable hydrolases with desired functions from hot terrestrial and marine environments. To this end, hot springs worldwide were mined, resulting in hundreds of environmental samples and thousands of enrichment cultures growing on polymeric substrates of industrial interest. Using high-throughput sequencing and bioinformatics, 15 hot spring metagenomes, as well as several sequenced isolate genomes and transcriptomes were obtained. To facilitate the discovery of novel hydrolases, the annotation platform Anastasia and a whole-cell bioreporter-based functional screening method were developed. Sequence-based screening and functional screening together resulted in about 100 potentially new hydrolases of which more than a dozen have been characterized comprehensively from a biochemical and structural perspective. The characterized hydrolases include thermostable carboxylesterases, enol lactonases, quorum sensing lactonases, gluconolactonases, epoxide hydrolases, and cellulases. Apart from these novel thermostable hydrolases, the project generated an enormous amount of samples and data, thereby allowing the future discovery of even more novel enzymes.