Discovery of two novel β-glucosidases from an Amazon soil metagenomic library

FEMS Microbiol Lett. 2014 Feb;351(2):147-55. doi: 10.1111/1574-6968.12332. Epub 2013 Dec 16.

Abstract

An Amazon soil microbial community metagenomic fosmid library was functionally screened for β-glucosidase activity. Contig analysis of positive clones revealed the presence of two ORFs encoding novel β-glucosidases, AmBGL17 and AmBGL18, from the GH3 and GH1 families, respectively. Both AmBGL17 and AmBGL18 were functionally identified as β-glucosidases. The enzymatic activity of AmBGL17 was further characterized. AmBGL17 was tested with different substrates and showed highest activity on pNPβG substrate with an optimum temperature of 45 °C and an optimum pH of 6. AmBGL17 showed a Vmax of 116 mM s(-1) and Km of 0.30 ± 0.017 mM. This is the first report of β-glucosidases from an Amazon soil microbial community using a metagenomic approach.

Keywords: GH1; GH3; beta-glucosidase; functional screen; metagenomic library.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Cellulases / chemistry
  • Cellulases / genetics
  • Cellulases / isolation & purification*
  • Cellulases / metabolism*
  • Enzyme Stability
  • Gene Library
  • Genetic Testing
  • Hydrogen-Ion Concentration
  • Metagenomics*
  • Molecular Sequence Data
  • Sequence Analysis, DNA
  • Soil Microbiology*
  • Substrate Specificity
  • Temperature

Substances

  • Cellulases

Associated data

  • GENBANK/KF433952
  • GENBANK/KF433953