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  • Purification and biochemical characterization of a specific beta-glucosidase from the digestive fluid of larvae of the palm weevil, Rhynchophorus palmarum.

Purification and biochemical characterization of a specific beta-glucosidase from the digestive fluid of larvae of the palm weevil, Rhynchophorus palmarum.

Journal of insect science (Online) (2009-07-21)
Désiré Yapi Assoi Yapi, Dago Gnakri, Sebastien Lamine Niamke, Lucien Patrice Kouame
ABSTRACT

A beta-glucosidase was purified from the digestive fluid of the palm weevil Rhynchophorus palmarum L. (Coleoptera: Curculionidae) by chromatography on anion-exchange, gel filtration, and hydrophobic interaction columns. The preparation was shown to be homogeneous on polyacrylamide gels, beta-glucosidase is a monomeric protein with a molecular weight of 58 kDa based on its mobility in SDS-PAGE and 60 kDa based on gel filtration. Maximal beta-glucosidase activity occurred at 55 degrees C and pH 5.0. The purified beta-glucosidase was stable at 37 degrees C and its pH stability was in the range of 5.0-6.0. The enzyme readily hydrolyzed p-nitrophenyl-beta-D-glucoside, cellobiose, cellodextrins and required strictly beta-gluco configuration for activity. It cleaved glucose-glucose beta-(1-4) linkages better than beta-(1-2), beta-(1-3) and beta-(1-6) linkages. The catalytic efficiency (K(cat)/K(M)) values for p-nitrophenyl-beta-D-glucoside and cellobiose were respectively 240.48 mM(-1)s(-1) and 134.80 mM(-1)s(-1). Beta-glucosidase was capable of catalysing transglucosylation reactions. The yield of glucosylation of 2-phenylethanol (20 %), catalysed by the beta-glucosidase in the presence of cellobiose as glucosyl donor, is lower than those reported previously with conventional sources of beta-glucosidases. In addition, the optimum pH is different for the hydrolysis (pH 5.0) and transglucosylation reactions (pH 6.6).

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Sophorose, ≥98% (HPLC, anhydrous)
Sigma-Aldrich
β-Gentiobiose, ≥85% (remainder primarily α-anomer)
Sigma-Aldrich
Xylobiose, ≥90% (HPLC)