Cloning, purification, and characterization of thermostable hypoxanthine-guanine phosphoribosyltransferase from Thermoanaerobacter tengcongensis

被引：9
作者：
Chen, Q
You, DL
Hua, MH
Gu, XC
Luo, M
Lu, SY [1 ]
机构：
[1] Peking Univ, Coll Life Sci, Struct Biol Lab, Beijing 100871, Peoples R China
[2] Univ Alabama Birmingham, Dept Microbiol, Birmingham, AL 35294 USA
来源：
PROTEIN EXPRESSION AND PURIFICATION | 2003年 / 32卷 / 02期
基金：
中国国家自然科学基金;
关键词：
hypoxanthine-guanine phosphoribosyltransferase; Thermoanaerobacter tengeongensis; enzyme kinetics; thermostable;
D O I：
10.1016/j.pep.2003.08.001
中图分类号：
Q5 [生物化学];
学科分类号：
071010 ; 081704 ;
摘要：
Hypoxanthine-guanine phosphoribosyltransferase (HGPRT, EC 2.4.2.8) from a newly characterized thermophile Thermoanaerobacter tengcongensis was expressed in Escherichia coli and purified. Analytical get filtration suggested that the enzyme exist as a homotetramer in solution. The optimal pH for the forward reaction was found to be 8.0 and the optimal temperature 70degreesC. The steady-state kinetic characteristics suggest that hypoxanthine is the most effective substrate. This enzyme showed a half-life of 75 min at 50degreesC and no apparent loss of activity after 3 months at 4degreesC. (C) 2003 Elsevier Inc. All rights reserved.
引用
收藏
页码：239 / 245
页数：7
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