AFFINITY-CHROMATOGRAPHY OF GLYCOENZYMES AND GLYCOPROTEINS ON CONCANAVALIN-A BEAD CELLULOSE

Concanavalin A immobilized on chlorotriazine bead cellulose was applie d to affinity purification of glycoenzymes and glycoproteins. Enzymes such as invertase from baker's yeast, endopolygalacturonase (Rohament P) and exopolygalacturonase from carrot roots, as well as extracellula r mannoproteins from the yeast Cryptococcus laurentii were examined. C hromatography was performed on minicolumns filled with Concanavalin A- triazine bead cellulose gel with the content of immobilized Concanaval in A within the range 1.2 - 8.2 mg per mt of gel. The specifically bou nd glycoenzymes or glycoproteins were eluted with a solution of the co rresponding counter-ligand alpha-methyl mannopyranoside. Individual de grees of purification, estimated from the measurements of specific act ivity of crude and purified glycoenzymes, were 14.5-fold for invertase , 93-fold for polygalacturonase and 3.9-fold for exopolygalacturonase. The yeast mannoprotein was isolated from the heteroglycoprotein fract ion. The purified mannoprotein contained mainly mannose, with traces o f glucose. The purification effect was verified by FPL-chromatography.