Ultrahigh-throughput discovery of promiscuous enzymes by picodroplet functional metagenomics.

ABSTRACT: Unculturable bacterial communities provide a rich source of biocatalysts, but their experimental discovery by functional metagenomics is difficult, because the odds are stacked against the experimentor. Here we demonstrate functional screening of a million-membered metagenomic library in microfluidic picolitre droplet compartments. Using bait substrates, new hydrolases for sulfate monoesters and phosphotriesters were identified, mostly based on promiscuous activities presumed not to be under selection pressure. Spanning three protein superfamilies, these break new ground in sequence space: promiscuity now connects enzymes with only distantly related sequences. Most hits could not have been predicted by sequence analysis, because the desired activities have never been ascribed to similar se

SUBMITTER: Colin PY 

PROVIDER: S-EPMC4686663 | biostudies-literature | 2015 Dec

REPOSITORIES: biostudies-literature