Activity screening of environmental metagenomic libraries reveals novel carboxylesterase families.
Activity screening of environmental metagenomic libraries reveals novel carboxylesterase families.
复制标题
DOI：
10.1038/srep44103
复制
发表时间：
2017-03-08
影响因子：
4.6
通讯作者：
Yakunin AF
中科院分区：
文献类型：
--
作者：
Popovic A;Hai T;Tchigvintsev A;Hajighasemi M;Nocek B;Khusnutdinova AN;Brown G;Glinos J;Flick R;Skarina T;Chernikova TN;Yim V;Brüls T;Paslier DL;Yakimov MM;Joachimiak A;Ferrer M;Golyshina OV;Savchenko A;Golyshin PN;Yakunin AF
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.
