Characterization of a novel beta-glucosidase-like activity from a soil metagenome.

We report the cloning of a novel beta-glucosidase-like gene by function-based screening of a metagenomic library from uncultured soil microorganisms. The gene was named bgllC and has an open reading frame of 1,443 base pairs. It encodes a 481 amino acid polypeptide with a predicted molecular mass of about 57.8 kDa. The deduced amino acid sequence did not show any homology with known beta-glucosidases. The putative beta-glucosidase gene was subcloned into the pETBlue-2 vector and overexpressed in E. coli Tuner (DE3) pLacI; the recombinant protein was purified to homogeneity. Functional characterization with a high performance liquid chromatography method demonstrated that the recombinant BgllC protein hydrolyzed D-glucosyl-beta-(l-4)-D-glucose to glucose. The maximum activity for BgllC protein occurred at pH 8.0 and 42 degrees C using p-nitrophenyl-beta-D-glucoside as the substrate. A CaCl(2) concentration of 1 mM was required for optimal activity. The putative beta-glucosidase had an apparent K(m) value of 0.19 mM, a V(max) value of 4.75 U/mg and a k (cat) value of 316.7/min under the optimal reaction conditions. The biochemical characterization of BgllC has enlarged our understanding of the novel enzymes that can be isolated from the soil metagenome.