title: Purification and properties of an amylopullulanase, a glucoamylase, and an alpha-glucosidase in the amylolytic enzyme system of Thermoanaerobacterium thermosaccharolyticum.
abstract: 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 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 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.
authors.affiliation: Institute for Microbiology, Technical University of Munich, Germany.
authors.forename: D
authors.initials: D
authors.lastname: Ganghofner
authors.forename: J
authors.initials: J
authors.lastname: Kellermann
authors.forename: W L
authors.initials: WL
authors.lastname: Staudenbauer
authors.forename: K
authors.initials: K
authors.lastname: Bronnenmeier
mesh_terms: D000595:Amino Acid Sequence; D005591:Chemical Fractionation; D002847:Chromatography, Agarose; D002851:Chromatography, High Pressure Liquid; D002852:Chromatography, Ion Exchange; D003013:Clostridium / Q000201:enzymology*; D004591:Electrophoresis, Polyacrylamide Gel; D005087:Glucan 1,4-alpha-Glucosidase / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism; D005936:Glucans / Q000378:metabolism; D005947:Glucose / Q000378:metabolism; D006026:Glycoside Hydrolases / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism; D006863:Hydrogen-Ion Concentration; D006868:Hydrolysis; D007525:Isoelectric Focusing; D008969:Molecular Sequence Data; D008970:Molecular Weight; D016415:Sequence Alignment; D013213:Starch / Q000378:metabolism; D013379:Substrate Specificity; D013696:Temperature; D014462:Ultrafiltration; D000520:alpha-Glucosidases / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism
chemical_list: D005936:Glucans; C009109:pullulan; D013213:Starch; D006026:Glycoside Hydrolases; D000520:alpha-Glucosidases; D005087:Glucan 1,4-alpha-Glucosidase; C100181:pullulanase; D005947:Glucose