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Isolation of thermostable phosphatase from the hyperthermophilic archaeon Thermococcus pacificus by immobilized metal affinity chromatography
Author(s) -
Bannikova Galina E.,
Varlamov Valery P.,
Miroshnichenko Margarita L.,
BonchOsmolovskaya Elizaveta A.
Publication year - 1998
Publication title -
iubmb life
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.132
H-Index - 113
eISSN - 1521-6551
pISSN - 1521-6543
DOI - 10.1080/15216549800201372
Subject(s) - size exclusion chromatography , affinity chromatography , chemistry , agarose , enzyme , phosphatase , pyrococcus furiosus , biochemistry , chromatography , gel permeation chromatography , protein subunit , archaea , organic chemistry , gene , polymer
Abstract Phosphatase was isolated from cells of the hyperthermophilic marine archaeon Thermococcus pacificus by a procedure including chromatography on Butyl‐Fractogel TSK‐650 and Ni2+‐iminodiacetic‐agarose. Enzyme activity is maximal at 90°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.