Department of Applied Chemistry, Graduate School of Engineering, Kyushu University, 744 Motooka, Fukuoka, Japan.
Biotechnol Lett. 2012 Nov;34(11):2055-60. doi: 10.1007/s10529-012-0998-0. Epub 2012 Jul 10.
Treatment of a hyperthermophilic enzyme, alkaline phosphatase from Pyrococcus furiosus (PfuAP), with EDTA completely deactivated PfuAP, indicating that the presence of one or more divalent metal ions is essential for its catalytic activity. Subsequent addition of various divalent metal ions to the apoprotein recovered the enzymatic activity and, in particular, the addition of Co(II) resulted in an over 50-fold increase in activity compared with PfuAP before EDTA treatment. Intriguingly, PfuAP with Co(II) exhibited weaker stability toward heat treatment, suggesting that Co(2+) destabilizes the tertiary structure of PfuAP at high temperature.
用 EDTA 处理嗜热酶、来自 Pyrococcus furiosus 的碱性磷酸酶（PfuAP）会使其完全失活，这表明一个或多个二价金属离子的存在对其催化活性是必需的。随后向脱辅基蛋白中添加各种二价金属离子可恢复酶活性，特别是添加 Co(II)可使酶活性比 EDTA 处理前的 PfuAP 增加超过 50 倍。有趣的是，与 Co(II)结合的 PfuAP 对热处理的稳定性较弱，表明 Co(2+)在高温下使 PfuAP 的三级结构不稳定。
