Single residues dictate the co-evolution of dual esterases: MCP hydrolases from the α/β hydrolase family
Single residues dictate the co-evolution of dual esterases: MCP hydrolases from the α/β hydrolase family
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DOI：
10.1042/bj20130552
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发表时间：
2013-08-15
影响因子：
4.1
通讯作者：
Ferrer, Manuel
中科院分区：
文献类型：
--
作者：
Alcaide, Maria;Tornes, Jesus;Ferrer, Manuel
C-C MCP（中间裂解产物）水解酶家族的几个成员表现出不同寻常的水解酯和MCP的能力（包括那些来自单环和双环芳烃）。尽管导致这种底物混杂的分子机制开始出现，但对这些复杂酶的全面了解还远远没有完成。在本文中，我们描述了通过基因组方法鉴定的六种不同的α / β水解酶，其中四种酶对广泛的底物具有前所未有的活性特征，包括对硝基苯基、卤化、脂肪酰基、芳基、甘油、肉桂基和碳水化合物酯、内酯、2-羟基-6-氧-6-苯六酸2,4-二烯酸酯和2-羟基-6-氧七酸2,4-二烯酸酯。通过结构分析和定点诱变，我们确定了三个残基(Ser(32), Val（130）和Trp(144))，它们决定了其中一个蛋白质CCSP0084不寻常的底物特异性。研究结果为MCP水解酶的比较催化模型、结构机理研究和生物技术应用开辟了新的研究途径。
Several members of the C-C MCP (meta-cleavage product) hydrolase family demonstrate an unusual ability to hydrolyse esters as well as the MCPs (including those from mono- and bi-cyclic aromatics). Although the molecular mechanisms responsible for such substrate promiscuity are starting to emerge, the full understanding of these complex enzymes is far from complete. In the present paper, we describe six distinct alpha/beta hydrolases identified through genomic approaches, four of which demonstrate the unprecedented characteristic of activity towards a broad spectrum of substrates, including p-nitrophenyl, halogenated, fatty acyl, aryl, glycerol, cinnamoyl and carbohydrate esters, lactones, 2-hydroxy-6-oxo-6-phenylhexa-2,4-dienoate and 2-hydroxy-6-oxohepta-2,4-dienoate. Using structural analysis and site-directed mutagenesis we have identified the three residues (Ser(32), Val(130) and Trp(144)) that determine the unusual substrate specificity of one of these proteins, CCSP0084. The results may open up new research avenues into comparative catalytic models, structural and mechanistic studies, and biotechnological applications of MCP hydrolases.