A cell wall-bound β-glucosidase from germinated rice:: Purification and properties

A large portion of beta-glucosidase (EC 3.2.1.21) in germinating rice seeds, which appears to be ionically bound to cell walls, can be solubilized with 1 M NaCl. Its activity increased more than eight-fold within five days of germination. It was purified to electrophoretic homogeneity from the extracts of germinated rice seeds by fractionation with (NH4)(2)SO4 followed by CM-Sepharose, Polybuffer exchanger 118, Concanavalin A-Sepharose and Bio-Gel P-100. The Mr of the purified enzyme, estimated by SDS-PAGE, was 56,000 and the isoelectric point was >10.0. Its N-terminal amino acid sequence (44 residues) exhibited high homology to those of P-glucosidases from other plants, such as barley and white clover. Its activity was optimal at pH 4.5 and 50 degrees, and it was strongly inhibited by glucono-1,5-lactone. The enzyme showed hydrolytic as well as transglycosylation activity towards (1 --> 3)-beta- and (1 --> 4)-beta-linked oligosaccharides with degree of polymerization of 2-4. The results suggest that the beta-glucosidase is probably involved not only in hydrolysis but also in modification of oligosaccharides in cell walls of germinating rice seeds. (C) 1998 Elsevier Science Ltd. All rights reserved.