title: Purification and characterization of a novel enzyme, maltooligosyl trehalose trehalohydrolase, from Arthrobacter sp. Q36.
abstract: A novel enzyme, maltooligosyl trehalose trehalohydrolase from Arthrobacter sp. Q36 was purified from a cell-free extract to an electrophoretically pure state by successive column chromatography on Sepabeads FP-DA13, Butyl-Toyopearl 650M, DEAE-Toyopearl 650S, and Toyopearl HW-55S. The enzyme specifically catalyzed the hydrolysis of the alpha-1,4-glucosidic linkage that bound the maltooligosyl and trehalose moieties of maltooligosyl trehalose. The Km of the enzyme for maltosyl trehalose, maltotriosyl trehalose, maltotetraosyl trehalose, and maltopentaosyl trehalose was 5.5 mM, 4.6 mM, 7.0 mM, and 4.2 mM, respectively. The enzyme had a molecular mass of 62,000 by SDS-polyacrylamide gel electrophoresis and a pI of 4.1 by gel isoelectrofocusing. The N-terminal amino acid of the enzyme was threonine. The enzyme showed the highest activity at pH 6.5 and 45 degrees C, and was stable from pH 5.0 to 10.0 and up to 45 degrees C. The activity was inhibited by Hg2+, Cu2+, Fe2+, and Zn2+.
authors.affiliation: Hayashibara Biochemical Laboratories, Inc., Okayama, Japan.
authors.forename: T
authors.initials: T
authors.lastname: Nakada
authors.forename: K
authors.initials: K
authors.lastname: Maruta
authors.forename: H
authors.initials: H
authors.lastname: Mitsuzumi
authors.forename: M
authors.initials: M
authors.lastname: Kubota
authors.forename: H
authors.initials: H
authors.lastname: Chaen
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; D001173:Arthrobacter / Q000201:enzymology*; D001426:Bacterial Proteins / Q000302:isolation & purification* / Q000378:metabolism*; D055598:Chemical Phenomena; D002627:Chemistry, Physical; D005959:Glucosidases / Q000302:isolation & purification* / Q000378:metabolism*; D008320:Maltose / Q000378:metabolism; D008969:Molecular Sequence Data; D008970:Molecular Weight; D013213:Starch / Q000378:metabolism; D013379:Substrate Specificity; D014199:Trehalose / Q000096:biosynthesis
chemical_list: D001426:Bacterial Proteins; D008320:Maltose; D013213:Starch; D014199:Trehalose; D005959:Glucosidases; C099156:maltooligosyl trehalose trehalohydrolase