Isolation of thermostable phosphatase from the hyperthermophilic archaeon Thermococcus pacificus by immobilized metal affinity chromatography.
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.
Bannikova
G E
GE
Bioengineering Center, Russian Academy of Sciences, Moscow, Russia.
Varlamov
V P
VP
Miroshnichenko
M L
ML
Bonch-Osmolovskaya
E A
EA
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*
D002413:Cations, Divalent; D003300:Copper; D009532:Nickel; D010744:Phosphoric Monoester Hydrolases