Activity screening of environmental metagenomic libraries reveals novel carboxylesterase families
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- Big_esterase_paper_Sci_Rep_4
Accepted author manuscript, 241 KB, PDF-document
License: CC BY Show license
- srep44103
Final published version, 2 MB, PDF-document
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DOI
- 10.1038/srep44103
Final published version
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 language | English |
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Article number | 44103 |
Journal | Scientific Reports |
Volume | 7 |
Issue number | 44103 |
Early online date | 8 Mar 2017 |
DOIs | |
State | E-pub ahead of print - 8 Mar 2017 |
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