Aii810, a Novel Cold-Adapted N-Acylhomoserine Lactonase Discovered in a Metagenome, Can Strongly Attenuate Pseudomonas aeruginosa Virulence Factors and Biofilm Formation.
Aii810, a Novel Cold-Adapted N-Acylhomoserine Lactonase Discovered in a Metagenome, Can Strongly Attenuate Pseudomonas aeruginosa Virulence Factors and Biofilm Formation.
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Aii810 是一种在宏基因组中发现的新型冷适应 N-酰基高丝氨酸内酯酶，可强烈减弱铜绿假单胞菌毒力因子和生物膜形成
DOI：
10.3389/fmicb.2017.01950
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发表时间：
2017
影响因子：
5.2
通讯作者：
Liu X
中科院分区：
文献类型：
--
作者：
Fan X;Liang M;Wang L;Chen R;Li H;Liu X
病原体铜绿假单胞菌使用群体感应（QS）来控制毒力和生物膜形成。酶促破坏群体感应是一种不依赖抗生素的有前途的抗感染治疗策略。本文首次从毛豆腐宏基因组中分离到一个编码N-酰基高丝氨酸内酯酶的新基因（aii 810）。Aii 810编码269个氨基酸的蛋白质，并在大肠杆菌BL 21（DE 3）中以可溶性形式表达。在20°C时酶活最高，0°C时酶活保持在76.5%以上，0-40°C时酶活保持在50%以上。最适pH为8.0。它在中性和微碱性条件下以及在40°C以下的温度下都是稳定的。该酶能水解多种对硝基苯酚酯，但最佳底物为乙酸对硝基苯酚酯。其kcat和Km值分别为347.7 S-1和205.1 μM。它有效地降解N-丁酰基-L-高丝氨酸内酯和N-（3-氧代十二烷酰基）-L-高丝氨酸内酯，分别超过72.3%和100%的水解率。此外，Aii 810强烈减弱铜绿假单胞菌毒力和生物膜形成。这种具有高抗QS活性的酶是所报道的最冷适应的N-酰基高丝氨酸内酯酶，这使其成为用作针对铜绿假单胞菌感染的治疗剂的有吸引力的酶。
The pathogen Pseudomonas aeruginosa uses quorum sensing (QS) to control virulence and biofilm formation. Enzymatic disruption of quorum sensing is a promising anti-infection therapeutic strategy that does not rely on antibiotics. Here, a novel gene (aii810) encoding an N-acylhomoserine lactonase was isolated from the Mao-tofu metagenome for the first time. Aii810 encoded a protein of 269 amino acids and was expressed in Escherichia coli BL21 (DE3) in soluble form. It showed the highest activity at 20°C, and it maintained 76.5% of activity at 0°C and more than 50% activity at 0–40°C. The optimal pH was 8.0. It was stable in both neutral and slightly alkaline conditions and at temperatures below 40°C. The enzyme hydrolyzed several ρ-nitrophenyl esters, but its best substrate was ρ-nitrophenyl acetate. Its kcat and Km values were 347.7 S-1 and 205.1 μM, respectively. It efficiently degraded N-butyryl-L-homoserine lactone and N-(3-oxododecanoyl)-L-homoserine lactone, exceeding hydrolysis rates of 72.3 and 100%, respectively. Moreover, Aii810 strongly attenuated P. aeruginosa virulence and biofilm formation. This enzyme with high anti-QS activity was the most cold-adapted N-acylhomoserine lactonase reported, which makes it an attractive enzyme for use as a therapeutic agent against P. aeruginosa infection.