title: A thermostable acid alpha-glucosidase from Tetrahymena thermophila: purification and characterization.
abstract: An acid alpha-glucosidase (EC 3.2.1.20) was purified to homogeneity from the culture medium of Tetrahymena thermophila CU 399. Its general molecular, catalytic and immunological properties were compared to those of the T. pyriformis W enzyme. The enzyme from T. thermophila was a 105-kD monomer and the N-terminus (25 amino acid residues) displayed some homology with that of T. pyriformis enzyme. The purified enzyme was most active at 56 degrees C and showed resistance to thermal inactivation. The acid alpha-glucosidase appears to have alpha-1,6-glucosidase as well as alpha-1,4-glucosidase activity. The Km values determined with p-nitrophenyl-alpha-glucopyranoside, maltose, isomaltose and glycogen were 0.7 mM, 2.5 mM, 28.5 mM and 18.5 mg/ml, respectively. The enzyme was antigenically distinct from T. pyriformis acid alpha-glucosidase.
authors.affiliation: Department of Biochemistry, Gifu University School of Medicine, Japan.
authors.forename: Y
authors.initials: Y
authors.lastname: Banno
authors.forename: N
authors.initials: N
authors.lastname: Sasaki
authors.forename: T
authors.initials: T
authors.lastname: Yoshino
authors.forename: J
authors.initials: J
authors.lastname: Mochizuki
authors.forename: H
authors.initials: H
authors.lastname: Hirata
authors.forename: Y
authors.initials: Y
authors.lastname: Nozawa
mesh_terms: D000595:Amino Acid Sequence; D000596:Amino Acids / Q000032:analysis; D000818:Animals; D000913:Antibodies, Protozoan / Q000276:immunology; D003429:Cross Reactions; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D008969:Molecular Sequence Data; D008970:Molecular Weight; D013379:Substrate Specificity; D013696:Temperature; D013768:Tetrahymena / Q000201:enzymology* / Q000276:immunology; D000520:alpha-Glucosidases / Q000032:analysis / Q000276:immunology / Q000302:isolation & purification* / Q000378:metabolism
chemical_list: D000596:Amino Acids; D000913:Antibodies, Protozoan; D000520:alpha-Glucosidases