Substrate specificity and kinetic properties of enzymes belonging to the hormone-sensitive lipase family: Comparison with non-lipolytic and lipolytic carboxylesterases
Substrate specificity and kinetic properties of enzymes belonging to the hormone-sensitive lipase family: Comparison with non-lipolytic and lipolytic carboxylesterases
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DOI：
10.1016/j.bbalip.2005.11.003
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发表时间：
2005-12-30
影响因子：
4.8
通讯作者：
Sarda, L
中科院分区：
文献类型：
--
作者：
Chahinian, H;Ben Ali, Y;Sarda, L
我们研究了HSL家族的四种羧酸酯酶水解三酰甘油、乙烯基酯和对硝基苯基丁酸酯的动力学，所述四种羧酸酯酶为重组人脂肪酶敏感脂肪酶（HSL）、来自酸热脂环酸芽孢杆菌的EST 2、来自闪烁古生球菌的AFEST和来自结核分枝杆菌的蛋白RV 1399 C。已经将HSL家族的酶的动力学性质与一系列脂解和非脂解羧酸酯酶的动力学性质进行了比较，所述脂解和非脂解羧酸酯酶包括人胰脂肪酶、豚鼠胰脂肪酶相关蛋白2、来自米黑毛霉和嗜热菌的脂肪酶、来自腐皮镰刀菌的角质酶、来自枯草芽孢杆菌的利帕、猪肝酯酶和来自尼日尔曲霉的酯酶A。结果表明，人HSL，与其他脂肪分解羧酸酯酶，是活性的短链酯和水解水不溶性的三辛酸甘油酯，月桂酸乙烯酯和橄榄油，而EST 2，AFEST，蛋白质RV 1399 C和非脂肪分解羧酸酯酶的行动是有限的短链底物的解决方案。脂解和非脂解羧酸酯酶可以通过它们各自的短链酯水解的K-0.5（表观K-m）值来区分。在脂解酶中，那些具有盖结构域的酶对三丁酸甘油酯、三辛酸甘油酯和橄榄油显示出更高的活性，这表明盖结构有助于酶与三酰基甘油的结合。通过具有盖结构域的脂解羧酸酯酶水解对硝基苯基丁酸酯的进展反应曲线显示潜伏期，这在人HSL、非脂解羧酸酯酶和缺乏盖结构的脂解酶如角质酶中未观察到。(c)2005 Elsevier B.V保留所有权利。
We have studied the kinetics of hydrolysis of triacylglycerols, vinyl esters and p-nitrophenyl butyrate by four carboxylesterases of the HSL family, namely recombinant human hormone-sensitive lipase (HSL), EST2 from Alicyclobacillus acidocaldarius, AFEST from Archeoglobus fulgidus, and protein RV1399C from Mycobacterium tuberculosis. The kinetic properties of enzymes of the HSL family have been compared to those of a series of lipolytic and non-lipolytic carboxylesterases including human pancreatic lipase, guinea pig pancreatic lipase related protein 2, lipases from Mucor miehei and Thermomyces lanuginosus, cutinase from Fusarium solani, LipA from Bacillus subtilis, porcine liver esterase and Esterase A from Aspergilus niger. Results indicate that human HSL, together with other lipolytic carboxylesterases, are active on short chain esters and hydrolyze water insoluble trioctanoin, vinyl laurate and olive oil, whereas the action of EST2, AFEST, protein RV1399C and non-lipolytic carboxylesterases is restricted to solutions of short chain substrates. Lipolytic and non-lipolytic carboxylesterases can be differentiated by their respective value of K-0.5 (apparent K-m) for the hydrolysis of short chain esters. Among lipolytic enzymes, those possessing a lid domain display higher activity on tributyrin, trioctanoin and olive oil suggesting, then, that the lid structure contributes to enzyme binding to triacylglycerols. Progress reaction curves of the hydrolysis of p-nitrophenyl butyrate by lipolytic carboxylesterases with lid domain show a latency phase which is not observed with human HSL, non-lipolytic carboxylesterases, and lipolytic enzymes devoid of a lid structure as cutinase. (c) 2005 Elsevier B.V All rights reserved.