title: Purification and some characteristics of phosphatase of a psychrophile.
abstract: The phosphatase of a psychrophile was purified by ammonium sulfate fractionation, and a sequence of chromatographies on DEAE-Cellulofine, butyl-Cellulofine, Sephacryl S-100, and Mono-Q columns. The purified enzyme preparation was found to be electrophoretically homogeneous on native- and SDS-PAGE, and its molecular mass was determined to be 38.4 kDa by MALDI-TOF mass spectrometry. Maximal activity was observed at 30 degrees C and pH 6.0. Furthermore, the activity of this enzyme at 0 and 5 degrees C was 27 and 28%, respectively, of that at 30 degrees C. The enzyme was stable in the pH range of 6.0 to 8.0 and up to 20 degrees C. The enzyme was affected by metal ions; the activity was enhanced by Mg2+ and Ca2+ ions, but depressed by Zn2+ ions. Analysis of the amino acid composition indicated that this phosphatase contains no S-S bond, and only a few prolyl residues necessary to retain the rigid structure of a protein molecule. The phosphatase shows typical features of a cold enzyme; high catalytic activity at low temperature and rapid inactivation at an intermediate temperature.
authors.affiliation: Department of Biofunctional Chemistry, Faculty of Agriculture, Kobe University, Hyogo.
authors.forename: H
authors.initials: H
authors.lastname: Tsuruta
authors.forename: S T
authors.initials: ST
authors.lastname: Tsuneta
authors.forename: Y
authors.initials: Y
authors.lastname: Ishida
authors.forename: K
authors.initials: K
authors.lastname: Watanabe
authors.forename: T
authors.initials: T
authors.lastname: Uno
authors.forename: Y
authors.initials: Y
authors.lastname: Aizono
mesh_terms: D000818:Animals; D000864:Antarctic Regions; D002118:Calcium / Q000494:pharmacology; D002413:Cations, Divalent / Q000494:pharmacology; D004789:Enzyme Activation / Q000187:drug effects; D004795:Enzyme Stability; D006088:Gram-Negative Aerobic Bacteria / Q000201:enzymology*; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D008274:Magnesium / Q000494:pharmacology; D010744:Phosphoric Monoester Hydrolases / Q000737:chemistry* / Q000302:isolation & purification* / Q000378:metabolism; D012758:Shellfish / Q000382:microbiology; D013696:Temperature; D015032:Zinc / Q000494:pharmacology
chemical_list: D002413:Cations, Divalent; D010744:Phosphoric Monoester Hydrolases; D008274:Magnesium; D015032:Zinc; D002118:Calcium