Stabilization of Taq DNA polymerase at high temperature by protein folding pathways from a hyperthermophilic archaeon, Pyrococcus furiosus.
Stabilization of Taq DNA polymerase at high temperature by protein folding pathways from a hyperthermophilic archaeon, Pyrococcus furiosus.
复制标题
DOI：
10.1002/bit.20781
复制
发表时间：
2006-01
影响因子：
3.8
通讯作者：
P. Laksanalamai;A. Pavlov;A. Slesarev;F. Robb
中科院分区：
文献类型：
--
作者：
P. Laksanalamai;A. Pavlov;A. Slesarev;F. Robb
强烈热球菌是一种最适生长温度为100 ℃的嗜热古菌，编码三种蛋白伴侣，即小热休克蛋白（sHsp）、前折叠蛋白（Pfd）和伴侣蛋白（Cpn）。在这项研究中，我们报告的被动伴侣sHsp和Pfd的激烈可以提高蛋白质的重折叠活性的ATP依赖的Cpn从同一超嗜热菌。Taq聚合酶的热稳定性显著提高的组合的P.furiosus伴侣，显示在升高的温度和热循环过程中持续的蛋白质折叠活性。基于这些结果，我们提出，在超嗜热古菌，P. furiosus的蛋白质折叠装置可以用来提高高温生物催化剂的耐久性和成本效益。
Pyrococcus furiosus, a hyperthermophilic archaeon growing optimally at 100 degrees C, encodes three protein chaperones, a small heat shock protein (sHsp), a prefoldin (Pfd), and a chaperonin (Cpn). In this study, we report that the passive chaperones sHsp and Pfd from P. furiosus can boost the protein refolding activity of the ATP-dependent Cpn from the same hyperthermophile. The thermo-stability of Taq polymerase was significantly improved by combinations of P. furiosus chaperones, showing ongoing protein folding activity at elevated temperatures and during thermal cycling. Based on these results, we propose that the protein folding apparatus in the hyperthermophilic archaeon, P. furiosus can be utilized to enhance the durability and cost effectiveness of high temperature biocatalysts.