title: Isolation of thermostable phosphatase from the hyperthermophilic archaeon Thermococcus pacificus by immobilized metal affinity chromatography.
abstract: Phosphatase was isolated from cells of the hyperthermophilic marine archaeon Thermococcus pacificus by a procedure including chromatography on Butyl-Fractogel TSK-650 and Ni(2+)-iminodiacetic-agarose. Enzyme activity is maximal at 90 degrees C, and the enzyme half-life time at this temperature is 1 h. The pH optimum of phosphatase activity is 6.0. Electrophoresis under denaturating conditions yielded a subunit molecular weight of 45 kDa. On gel-filtration on Sephacryl S-300 HR three peak corresponding to 295, 85 and 45 kDa were observed, suggesting that the enzyme is a homohexamer.
authors.affiliation: Bioengineering Center, Russian Academy of Sciences, Moscow, Russia.
authors.forename: G E
authors.initials: GE
authors.lastname: Bannikova
authors.forename: V P
authors.initials: VP
authors.lastname: Varlamov
authors.forename: M L
authors.initials: ML
authors.lastname: Miroshnichenko
authors.forename: E A
authors.initials: EA
authors.lastname: Bonch-Osmolovskaya
mesh_terms: D002413:Cations, Divalent; D002846:Chromatography, Affinity / Q000379:methods*; D003300:Copper; D004795:Enzyme Stability; D006358:Hot Temperature; D006863:Hydrogen-Ion Concentration; D008970:Molecular Weight; D009532:Nickel; D010744:Phosphoric Monoester Hydrolases / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism; D019712:Thermococcus / Q000201:enzymology*
chemical_list: D002413:Cations, Divalent; D003300:Copper; D009532:Nickel; D010744:Phosphoric Monoester Hydrolases