Purification and characterisation of a beta-glucosidase abundantly expressed in ripe sweet cherry (Prunus avium L.) fruit

A beta -glucosidase (beta -D-glucoside glucohydrolase, EC 3.2.1.21) was pur ified to homogeneity from ripe fruits of sweet cherry (Prunus avium L.) by ammonium sulphate precipitation. ion exchange and size exclusion chromatogr aphy. The enzyme is a monomer with a molecular mass of similar to 68 kDa an d an acidic isoelectric point. N-terminal sequence analysis indicated that sweet cherry beta -glucosidase is related to other plant cyanogenic beta -g lucosidases. Substrate specificity studies revealed that the enzyme is able to attack and hydrolyse several synthetic substrates and total cell walls purified from ripe fruit. Biochemical and immunolocalisation studies showed that sweet cherry beta -glucosidases are mainly localised in the cytosol a nd in the apoplast, at the unripe stage of ripening; in ripe fruit it is al so associated with cell wall. (C) 2001 Elsevier Science Ireland Ltd. All ri ghts reserved.