AidB, a Novel Thermostable N-Acylhomoserine Lactonase from the Bacterium Bosea sp.
AidB, a Novel Thermostable N-Acylhomoserine Lactonase from the Bacterium Bosea sp.
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AidB，一种来自 Bosea sp. 的新型热稳定 N-酰基高丝氨酸内酯酶。
DOI：
10.1128/aem.02065-19
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发表时间：
2019
影响因子：
4.4
通讯作者：
Zhang Li-Qun
中科院分区：
文献类型：
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作者：
Zhang Jun-Wei;Xuan Chen-Guang;Lu Can-Hua;Guo Song;Yu Jin-Feng;Asif Muhammad;Jiang Wen-Jun;Zhou Zhi-Gang;Luo Zhao-Qing;Zhang Li-Qun
许多革兰氏阴性菌利用n -酰基高丝氨酸内酯（AHLs）作为群体感应（QS）信号分子，以密度依赖的方式调节毒力表达。通过ahl酶灭活的群体猝灭（QQ）是一种很有前景的减少细菌感染和耐药的策略。本文从土壤细菌boseasp中鉴定出一种耐热AHL内酯酶（AidB），该酶可以降解不同的AHL，无论是否在C-3位置取代羰基或羟基。应变F3-2。超高效液相色谱分析表明，AidB是一种AHL内酯酶，可水解高丝氨酸内酯（HSL）环的酯键。AidB在30 ~ 80℃范围内具有热稳定性，在60℃预孵育30 min后活性达到最大。AidB的最适温度为60℃，在4℃或室温双蒸馏水（ddH2O）中均能稳定保存。AidB同源物仅在α变形菌门的根瘤菌门和红螺旋菌门中发现。产自农杆菌的AidB和多医院根瘤菌的AidB（与AidB的氨基酸同源性分别为50.6%和52.8%）也表现出耐热性AHL降解活性。将表达AidB的质粒引入细菌后，发现AidB能降低铜绿假单胞菌PAO1的产花青素量，并能降低胡萝卜素Pectobacterium carotovorum subsp.cartovorumz3 -3的致病性，提示AidB是一种潜在的降解ahl的治疗药物。在许多细菌病原体中，以ahl为信号的群体感应系统是一种重要的毒力调节剂，也是抗感染药物的一个有吸引力的靶点。在这项工作中，我们从土壤细菌菌株Boseasp中鉴定出一种新的AHL内酯酶AidB。F3-2。aid的表达可促进铜绿假单胞菌和胡萝卜乳杆菌中AHL信号和qs依赖性毒力因子的产生。AidB具有ahl降解活性的同源物仅在阿尔法变形菌属的几个属中被发现。值得注意的是，AidB是一种耐热酶，在80°C下处理30 min后仍保持其催化活性，在4°C和室温下均表现出可靠的储存稳定性。这些特性可能使其更适合实际应用。
Many Gram-negative bacteria employN-acylhomoserine lactones (AHLs) as quorum-sensing (QS) signal molecules to regulate virulence expression in a density-dependent manner. Quorum quenching (QQ) via enzymatic inactivation of AHLs is a promising strategy to reduce bacterial infections and drug resistance. Herein, a thermostable AHL lactonase (AidB), which could degrade different AHLs, with or without a substitution of carbonyl or hydroxyl at the C-3 position, was identified from the soil bacteriumBoseasp. strain F3-2. Ultrahigh-performance liquid chromatography analysis demonstrated that AidB is an AHL lactonase that hydrolyzes the ester bond of the homoserine lactone (HSL) ring. AidB was thermostable in the range 30 to 80°C and showed maximum activity after preincubation at 60°C for 30 min. The optimum temperature of AidB was 60°C, and the enzyme could be stably stored in double-distilled water (ddH2O) at 4°C or room temperature. AidB homologs were found only inRhizobialesandRhodospirillalesof theAlphaproteobacteria. AidB from Agrobacterium tumefaciens and AidB from Rhizobium multihospitium (with amino acid identities of 50.6% and 52.8% to AidB, respectively) also showed thermostable AHL degradation activity. When introduced into bacteria, plasmid-expressed AidB attenuated pyocyanin production by Pseudomonas aeruginosa PAO1 and the pathogenicity of Pectobacterium carotovorum subsp.carotovorumZ3-3, suggesting that AidB is a potential therapeutic agent by degrading AHLs.IMPORTANCEA quorum-sensing system using AHLs as the signal in many bacterial pathogens is a critical virulence regulator and an attractive target for anti-infective drugs. In this work, we identified a novel AHL lactonase, AidB, from a soil bacterial strain,Boseasp. F3-2. The expression ofaidBreduced the production of AHL signals and QS-dependent virulence factors in Pseudomonas aeruginosa and Pectobacterium carotovorum. The homologs of AidB with AHL-degrading activities were found only in several genera belonging to theAlphaproteobacteria. Remarkably, AidB is a thermostable enzyme that retained its catalytic activity after treatment at 80°C for 30 min and exhibits reliable storage stability at both 4°C and room temperature. These properties might make it more suitable for practical application.