Activity screening of environmental metagenomic libraries reveals novel carboxylesterase families

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  • Anna Popovic
    University of Toronto, Canada
  • Tran Hai
  • Anatoly Tchigvintsev
    University of Toronto, Canada
  • Mahbod Hajighasemi
    University of Toronto, Canada
  • Boguslaw Nocek
    Argonne National Laboratory, Argonne, IL.
  • Anna N. Khusnutdinova
    University of Toronto, Canada
  • Greg Brown
    University of Toronto, Canada
  • Julia Glinos
    University of Toronto, Canada
  • Robert Flick
    University of Toronto, Canada
  • Tatiana Skarina
    University of Toronto, Canada
  • Tatyana Chernikova
  • Veronica Yim
    University of Toronto, Canada
  • Thomas Bruls
    Centre National de la Recherche Scientifique (CNRS)
  • Denis Le Paslier
    Centre National de la Recherche Scientifique (CNRS)
  • Michail M. Yakimov
    Institute for Coastal Marine Environment
  • Andrzej Joachimiak
    Argonne National Laboratory, Argonne, Illinois
  • Manuel Ferrer
    Institute of Catalysis, Madrid, Spain
  • Olga Golyshina
  • Alexei Savchenko
    University of Toronto, Canada
  • Peter Golyshin
  • Alexander F. Yakunin
    University of Toronto, Canada
Metagenomics has made accessible an enormous reserve of global biochemical diversity. 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. We have 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.
Original languageEnglish
Article number44103
JournalScientific Reports
Volume7
Issue number44103
Early online date8 Mar 2017
DOIs
StateE-pub ahead of print - 8 Mar 2017

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