A 1,4-β-glucan glucanohydrolase from the cellulolytic fungus Trichoderma viride QM 9414. Purification, characterization and preparation of an immunoadsorbent for the enzyme

Abstract

A 1,4-.beta.-glucan glucanohydrolase (EC 3.2.1.4) was isolated from culture filtrates of the fungus T. viride QM 9414 by molecular-sieve chromatography on Bio-Gel P-30, ion-exchange chromatography on DEAE-Sephadex A-50 and isoelectric focusing in a density gradient. Polyacrylamide-gel electrophoresis at 2 different pH values, analytical isoelectric focusing in a polyacrylamide-gel slab and molecular-sieve chromatography of the reduced and alkylated enzyme in a denaturing medium indicated homogeneous protein. The enzyme has a MW of 51,000 and is not a glycoprotein. The pI [isoelectric point] 4.66 at 23.degree. C. Antiserum against the purified enzyme was prepared and the amount of enzyme in the original filtrate was determined by rocket immunoelectrophoresis to be about 50 mg/l. An immunoadsorbent made from CNBr-activated Sepharose 4B and antiserum affords a rapid and highly specific purification of the enzyme.