title: Purification and characterization of thermostable maltooligosyl trehalose trehalohydrolase from the thermoacidophilic archaebacterium Sulfolobus acidocaldarius.
abstract: A thermostable maltooligosyl trehalose trehalohydrolase from the thermoacidophilic archaebacterium Sulfolobus acidocaldarius ATCC 33909 was purified from a cell-free extract to an electrophoretically pure state by successive column chromatographies on Sepabeads FP-DA13, Butyl-Toyopearl 650M, DEAE-Toyopearl 650S, Toyopearl HW-55S and Ultrogel AcA44. The enzyme had a molecular mass of 59,000 by SDS-polyacrylamide gel electrophoresis and a pI of 6.1 by gel isoelectrofocusing. The N-terminal amino acid of the enzyme was methionine. The enzyme showed the highest activity from pH 5.5 to 6.0 and at 75 degrees C, and was stable from pH 5.5 to 9.5 and up to 85 degrees C. The activity was inhibited by Hg2+, Cu2+, Fe2+, Pb2+, and Zn2+. The Km values of the enzyme for maltosyl trehalose, maltotriosyl trehalose, maltotetraosyl trehalose, and maltopentaosyl trehalose were 16.7 mM, 2.7 mM, 3.7 mM, and 4.9 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: D055598:Chemical Phenomena; D002627:Chemistry, Physical; D005959:Glucosidases / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism*; D007700:Kinetics; D013379:Substrate Specificity; D016944:Sulfolobus / Q000201:enzymology*; D014199:Trehalose / Q000378:metabolism
chemical_list: D014199:Trehalose; D005959:Glucosidases; C099156:maltooligosyl trehalose trehalohydrolase