通过固定化金属亲和色谱法从嗜热古菌太平洋栖热球菌中分离热稳定磷酸酶。

Isolation of thermostable phosphatase from the hyperthermophilic archaeon Thermococcus pacificus by immobilized metal affinity chromatography.

作者信息

Bannikova G E, Varlamov V P, Miroshnichenko M L, Bonch-Osmolovskaya E A

机构信息

Bioengineering Center, Russian Academy of Sciences, Moscow, Russia.

出版信息

Biochem Mol Biol Int. 1998 Feb;44(2):363-70. doi: 10.1080/15216549800201372.

DOI:10.1080/15216549800201372

PMID:9530518

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.

摘要

通过包括在丁基-Fractogel TSK-650和镍（2+）-亚氨基二乙酸琼脂糖上进行色谱分离的方法，从嗜热海洋古菌太平洋嗜热栖热菌的细胞中分离出磷酸酶。该酶活性在90℃时最大，在此温度下酶的半衰期为1小时。磷酸酶活性的最适pH为6.0。变性条件下的电泳显示亚基分子量为45 kDa。在Sephacryl S-300 HR上进行凝胶过滤时，观察到对应于295、85和45 kDa的三个峰，表明该酶是同型六聚体。