TY - JOUR
T1 - Discovering novel hydrolases from hot environments
AU - Wohlgemuth, Roland
AU - Littlechild, Jennifer
AU - Monti, Daniela
AU - Schnorr, Kirk
AU - van Rossum, Teunke
AU - Siebers, Bettina
AU - Menzel, Peter
AU - Kublanov, Ilya V.
AU - Rike, Anne Gunn
AU - Skretas, Georgios
AU - Szabo, Zalan
AU - Peng, Xu
AU - Young, Mark J.
PY - 2018/12
Y1 - 2018/12
N2 - 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.
AB - 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.
KW - Biocatalysis
KW - Enrichment
KW - Enzyme characterization
KW - Enzyme screening
KW - Gene expression
KW - Genomics
KW - Hydrolases
KW - Metagenomics
KW - Sequencing
KW - Thermophiles
U2 - 10.1016/j.biotechadv.2018.09.004
DO - 10.1016/j.biotechadv.2018.09.004
M3 - Journal article
C2 - 30266344
AN - SCOPUS:85056190720
SN - 0734-9750
VL - 36
SP - 2077
EP - 2100
JO - Biotechnology Advances
JF - Biotechnology Advances
IS - 8
ER -