Activity screening of environmental metagenomic libraries reveals novel carboxylesterase families

Popovic, Ana; Hai, Tran; Tchigvintsev, Anatoly; Hajighasemi, Mahbod; Nocek, Boguslaw; Khusnutdinova, Anna N.; Brown, Greg; Glinos, Julia; Flick, Robert; Skarina, Tatiana; Chernikova, Tatyana N.; Yim, Veronica; Brüls, Thomas; Paslier, Denis Le; Yakimov, Michail M.; Joachimiak, Andrzej; Ferrer, Manuel; Golyshina, Olga V.; Savchenko, Alexei; Golyshin, Peter N.; Yakunin, Alexander F.

doi:10.1038/srep44103

Title: Activity screening of environmental metagenomic libraries reveals novel carboxylesterase families

Journal Article · Tue Mar 07 23:00:00 EST 2017 · Scientific Reports

DOI: https://doi.org/10.1038/srep44103 · OSTI ID:1393927

Popovic, Ana ^[1]; Hai, Tran ^[2]; Tchigvintsev, Anatoly ^[1]; Hajighasemi, Mahbod ^[1]; Nocek, Boguslaw ^[3]; Khusnutdinova, Anna N. ^[1]; Brown, Greg ^[1]; Glinos, Julia ^[1]; Flick, Robert ^[1]; Skarina, Tatiana ^[1]; Chernikova, Tatyana N. ^[2]; Yim, Veronica ^[1]; Brüls, Thomas ^[4]; Paslier, Denis Le ^[5]; Yakimov, Michail M. ^[6]; Joachimiak, Andrzej ^[3]; Ferrer, Manuel ^[7]; Golyshina, Olga V. ^[2]; Savchenko, Alexei ^[1]; Golyshin, Peter N. ^[2] more »

Univ. of Toronto, ON (Canada). Dept. of Chemical Engineering and Applied Chemistry
Bangor Univ., Gwynedd (United Kingdom). School of Biological Sciences
Argonne National Lab. (ANL), Argonne, IL (United States). Midwest Center for Structural Genomics, Structural Biology Center
Univ. of Evry Val d'Essonne (UEVE) (France). National Center of Scientific Research (CNRS)
Univ. of Evry Val d'Essonne (UEVE) (France). National Center for Scientific Research (CNRS)
Inst. for Coastal Marine Environment, Messina (Italy)
Spanish National Research Council (CSIC), Madrid (Spain). Inst. of Catalysis

Metagenomics has made accessible an enormous reserve of global biochemical diversity. In order to tap into this vast resource of novel enzymes, we have screened over one million clones from metagenome DNA libraries derived from sixteen different environments for carboxylesterase activity and identified 714 positive hits. Here, we validated the esterase activity of 80 selected genes, which belong to 17 different protein families including unknown and cyclase-like proteins. Three metagenomic enzymes exhibited lipase activity, and seven proteins showed polyester depolymerization activity against polylactic acid and polycaprolactone. Detailed biochemical characterization of four new enzymes revealed their substrate preference, whereas their catalytic residues were identified using site-directed mutagenesis. The crystal structure of the metal-ion dependent esterase MGS0169 from the amidohydrolase superfamily revealed a novel active site with a bound unknown ligand. Thus, activity-centered metagenomics has revealed diverse enzymes and novel families of microbial carboxylesterases, whose activity could not have been predicted using bioinformatics tools.

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Research Organization:: Argonne National Lab. (ANL), Argonne, IL (United States)

Sponsoring Organization:: USDOE Office of Science (SC), Biological and Environmental Research (BER) (SC-23); Genome Canada; Ontario Genomics Institute; Ontario Research Fund; Natural Sciences and Engineering Research Council of Canada (NSERC); Spanish Ministerio de Economia y Competitividad (MINECO); German Federal Ministry of Education and Research (BMBF); Biotechnology and Biological Sciences Research Council (BBSRC); European Regional Development Fund (ERDF)

Grant/Contract Number:: AC02-06CH11357

OSTI ID:: 1393927

Journal Information:: Scientific Reports, Journal Name: Scientific Reports Vol. 7; ISSN 2045-2322

Publisher:: Nature Publishing GroupCopyright Statement

Country of Publication:: United States

Language:: English

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