Glycoprotein Nature of Glycosidases from Leaves of Pisum sativum L

-Mannosidase, ß-N-acetylglucosaminidase, - and ß-galactosidaseand ß-glucosidase were partially purified from leavesof Pisum sativum by ammonium sulphate fractionation and columnchromatography on DEAE-Sephadex A-50 and hydroxylapatite. Atleast two molecular forms of each enzyme were resolved by thesetechniques except for ß-glucosidase of which onlyone form was resolved. Except for one form of -galactosidase,all of the glycosidases thus purified were completely boundby Sepharose-linked Concanavalin A. The binding was stronglyinhibited by cr-methyl-D-mannoside and no binding to Sepharose-6-Boccurred indicating that these glycosidases contain mannose-richoligosaccharides. The glycoprotein nature of -mannosidase, ß-galactosidaseand ß-glucosidase was further demonstrated by chromatographyon phenylboronate agarose columns. The differences in the concentrationof cr-methyl-D-mannoside and sorbitol required to elute thevarious glycosidases from Sepharose-linked Con A and phenylboronateagarose, respectively, suggested that these enzymes are glycosylatedto various degrees or that structural variation in their carbohydratemoieties occur. This is the first demonstration that glycosylationof several glycosidases present in a single plant species isapparently a generalized feature of these enzymes. Key words: Pisum sativum, Glycosidase, Glycoprotein