title: Purification and characterization of thermostable maltooligosyl trehalose synthase from the thermoacidophilic archaebacterium Sulfolobus acidocaldarius.
abstract: A thermostable maltooligosyl trehalose synthase was purified from a cell-free extract of the thermoacidophilic archaebacterium Sulfolobus acidocaldarius ATCC 33909 to an electrophoretically homogeneous state by successive column chromatography on Sepabeads FP-DA13, Butyl-Toyopearl 650M, DEAE-Toyopearl 650S, Ultrogel AcA44, and Mono Q. The enzyme had a molecular mass of 74,000 by SDS-polyacrylamide gel electrophoresis and a pI of 5.9 by gel isoelectrofocusing. The N-terminal amino acid of the enzyme was methionine. The enzyme showed the highest activity from pH 5.0 to 5.5 and at 75 degrees C, and was stable from pH 4.5 to 9.5 and up to 85 degrees C. The enzyme activity was inhibited by Hg2+ and Cu2+. The Kms of the enzyme for maltotetraose, maltopentaose, maltohexaose, maltoheptaose, and short chain amylose (DP 18) were 41.5 mM, 7.1 mM, 5.7 mM, 1.4 mM, and 0.6 mM, respectively.
authors.affiliation: Hayashibara Biochemical Laboratories, Inc., Okayama, Japan.
authors.forename: T
authors.initials: T
authors.lastname: Nakada
authors.forename: S
authors.initials: S
authors.lastname: Ikegami
authors.forename: H
authors.initials: H
authors.lastname: Chaen
authors.forename: M
authors.initials: M
authors.lastname: Kubota
authors.forename: S
authors.initials: S
authors.lastname: Fukuda
authors.forename: T
authors.initials: T
authors.lastname: Sugimoto
authors.forename: M
authors.initials: M
authors.lastname: Kurimoto
authors.forename: Y
authors.initials: Y
authors.lastname: Tsujisaka
mesh_terms: D000595:Amino Acid Sequence; D055598:Chemical Phenomena; D002627:Chemistry, Physical; D004795:Enzyme Stability; D005964:Glucosyltransferases / Q000378:metabolism*; D006358:Hot Temperature; D007700:Kinetics; D008969:Molecular Sequence Data; D009844:Oligosaccharides / Q000378:metabolism*; D016944:Sulfolobus / Q000201:enzymology*; D014199:Trehalose / Q000378:metabolism*
chemical_list: D009844:Oligosaccharides; C021705:maltooligosaccharides; D014199:Trehalose; D005964:Glucosyltransferases; C099084:maltooligosyl trehalose synthase