Structural and Biochemical Characterization of AaL, a Quorum Quenching Lactonase with Unusual Kinetic Properties.
Structural and Biochemical Characterization of AaL, a Quorum Quenching Lactonase with Unusual Kinetic Properties.
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DOI：
10.1038/s41598-018-28988-5
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发表时间：
2018-07-26
影响因子：
4.6
通讯作者：
Elias M
中科院分区：
文献类型：
--
作者：
Bergonzi C;Schwab M;Naik T;Daudé D;Chabrière E;Elias M
群体淬灭内酯酶是能够通过其酶促降解破坏基于酰基高丝氨酸内酯（阿勒）的细菌信号传导的酶。特别是，内酯酶因此被证明可以抑制依赖于这些化学物质的细菌行为，例如生物膜的形成或毒力因子的表达。在这里，我们的特点，生化和结构的一个新的代表性的金属-β-内酰胺酶超家族，命名为AaL，是从嗜热嗜酸菌Alicyclobacillusacidoterrestris分离。AaL是一种有效的群体淬灭酶，如其抑制鲍曼不动杆菌生物膜形成的能力所证明的。动力学研究表明，AaL是一种高效且广谱的酶，能够以高速率（kcat/KM > 105 M-1.s-1）水解多种内酯。此外，AaL显示出异常低的KM值，范围为10至80 µM。结合磷酸，甘油和C6-阿勒的AaL结构的分析揭示了一个独特的疏水补丁（W26，F87和I237），参与底物结合，可能占酶的高特异性。识别特异性决定因素将有助于开发高度特异性的群体淬灭酶作为潜在的治疗方法。
Quorum quenching lactonases are enzymes that are capable of disrupting bacterial signaling based on acyl homoserine lactones (AHL) via their enzymatic degradation. In particular, lactonases have therefore been demonstrated to inhibit bacterial behaviors that depend on these chemicals, such as the formation of biofilms or the expression of virulence factors. Here we characterized biochemically and structurally a novel representative from the metallo-β-lactamase superfamily, named AaL that was isolated from the thermoacidophilic bacterium Alicyclobacillus acidoterrestris. AaL is a potent quorum quenching enzyme as demonstrated by its ability to inhibit the biofilm formation of Acinetobacter baumannii. Kinetic studies demonstrate that AaL is both a proficient and a broad spectrum enzyme, being capable of hydrolyzing a wide range of lactones with high rates (kcat/KM > 105 M−1.s−1). Additionally, AaL exhibits unusually low KM values, ranging from 10 to 80 µM. Analysis of AaL structures bound to phosphate, glycerol, and C6-AHL reveals a unique hydrophobic patch (W26, F87 and I237), involved in substrate binding, possibly accounting for the enzyme’s high specificity. Identifying the specificity determinants will aid the development of highly specific quorum quenching enzymes as potential therapeutics.
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