ISOLATION, PURIFICATION, AND PROPERTIES OF CELLOBIOHYDROLASE FROM TRICHODERMA-LONGIBRACHIATUM

A scheme of isolation and purification of cellobiohydrolase from Trichoderma longibrachiatum is proposed, based on preliminary group purification by precipitation with ammonium sulfate and ion exchange chromatography on DEAE-Spherone, followed by two operations of ion exchange chromatography on a column of Mono Q. This method permitted the production of an enzyme homogeneous according to the data of analytical isoelectrofocusing and electrophoresis in a polyacrylamide gel gradient. Cellobiohydrolase has a molecular weight of 65,000 D according to the data of electrophoresis, 54,000 D according to the data of high-performance gel filtration; an isoelectric point of 3.7; and it contains 1.6% carbohydrates. The distribution constant of the enzyme on hydrocellulose is 0.4 liter/g. The activities of cellobiohydrolase toward carbomethylcellulose, beta-glucan, amorphous cellulose, microcrystalline cellulose, and methylumbelliferyl-beta-D-cellobioside were determined.