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

• Published in: Bioscience, biotechnology, and biochemistry Vol. 62; no. 2; pp. 302 - 308
• Main Authors: Ganghofner, D. (Technical Univ., of Munich, Munchen (Germany)), Kellermann, J, Staudenbauer, W.L, Bronnenmeier, K
• Format: Journal Article
• Language: English
• Published: Tokyo Japan Society for Bioscience, Biotechnology, and Agrochemistry 01.02.1998; Japan Society for Bioscience Biotechnology and Agrochemistry
• Subjects: ALFA GLUCOSIDASA; ALPHA GLUCOSIDASE; alpha-Glucosidases - chemistry; alpha-Glucosidases - isolation & purification; alpha-Glucosidases - metabolism; Amino Acid Sequence; amylopullulanase; BACTERIA; Bacteriology; Biological and medical sciences; Biology of microorganisms of confirmed or potential industrial interest; Biotechnology; Chemical Fractionation; Chromatography, Agarose; Chromatography, High Pressure Liquid; Chromatography, Ion Exchange; Clostridium - enzymology; Electrophoresis, Polyacrylamide Gel; Fundamental and applied biological sciences. Psychology; Glucan 1,4-alpha-Glucosidase - chemistry; Glucan 1,4-alpha-Glucosidase - isolation & purification; Glucan 1,4-alpha-Glucosidase - metabolism; Glucans - metabolism; GLUCOAMILASA; GLUCOAMYLASE; Glucose - metabolism; Glycoside Hydrolases - chemistry; Glycoside Hydrolases - isolation & purification; Glycoside Hydrolases - metabolism; Hydrogen-Ion Concentration; Hydrolysis; Isoelectric Focusing; Metabolism. Enzymes; Microbiology; Mission oriented research; Molecular Sequence Data; Molecular Weight; Physiology and metabolism; PURIFICACION; PURIFICATION; RESISTANCE A LA TEMPERATURE; RESISTENCIA A LA TEMPERATURA; Sequence Alignment; Starch - metabolism; Substrate Specificity; Temperature; TEMPERATURE RESISTANCE; Thermoanaerobacterium thermosaccharolyticum; thermophilic enzymes; Ultrafiltration; α-glucosidase; Glucan 1,4-α-glucosidase; Temperature; Purification; Enzyme; Environmental factor; α-Dextrin endo-1,6-α-glucosidase; Thermoanaerobacterium thermosaccharolyticum; Enzymatic activity; α-Glucosidase; Glycosidases; Substrate specificity; pH; Bacteria; Hydrolases; O-Glycosidases; Amylolytic enzyme; Physicochemical properties
• ISSN: 0916-8451; 1347-6947
• DOI: 10.1271/bbb.62.302

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 alpha-glucosidase. The enzymes were purified to homogeneity and their physical and catalytic properties were studied. The pullulanase, which cleaved both alpha-1,4- and alpha-1,6-glucosidic bonds, was an amuylopullulanase 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 alpha-glucosidase was identified as an isomaltase belonging to a group of structurally related exo-alpha-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 alpha-glucosidase assisting in the cleavage of alpha-1,6-glucosidic bonds