Purification and properties of an amylopullulanase, a glucoamylase, and an α-glucosidase in the amylolytic enzyme system of thermoanaerobacterium thermosaccharolyticum

Thermoanaerobic bacteria are of considerable interest as producers of thermostable amylolytic enzymes. The soluble amylolytic enzyme system of Thermoanaerobacterium thermosaccharolyticum DSM 571 was fractionated into a pullulanase, a glucoamylase, and an α-glucosidase. The enzymes were purified to homogeneity and their physical and catalytic properties were studied. The pullulanase, which cleaved both α-1,4- and α-1,6-glucosidic bonds, was an amylopullulanase closely related to similar enzymes from other thermoanaerobic bacteria. Partial amino acid sequences of the glucoamylase were identical with the corresponding sequences deduced from the cga gene encoding the glucoamylase from Clostridium sp. strain G0005. The α-glucosidase was identified as an isomaltase belonging to a group of structurally related exo-α-1,4-glucosidases and oligo-1,6-glucosidases from bacilli. Comparison of enzyme activities indicated that the glucoamylase had the major amylolytic activity of T. thermosaccharolyticum, with amylopullulanase and α-glucosidase assisting in the cleavage of α-1,6-glucosidic bonds.