LC-MS/MS检测水产品源致病菌和腐败菌群体感应AHLs信号分子  

Detection of N-acylated-L-homoserine lactones of quorum sensing secreted by pathogen and spoilage bacteria from aquatic product using liquid chromatography-mass spectrometry /mass spectrometry

作　　者：黄旭镇 朱军莉[1] 赵二科 梁新乐[1] 

机构地区：[1]浙江工商大学食品与生物工程学院,浙江杭州310018

出　　处：《水产学报》2014年第7期1040-1046,共7页Journal of Fisheries of China

基　　金：国家自然科学基金(31271954;31171745);国家科技支撑计划(2012BAD29B06);浙江省科技厅项目(2012C22049);浙江工商大学校级研究生科技创新项目(1110XJ1513125)

摘　　要：为分析水产品源致病菌和腐败菌中群体感应信号分子AHLs的含量和种类,实验建立液相色谱-串联质谱(LC-MS/MS)同时检测细菌群体感应11种AHLs类信号分子的技术,并与生物报告菌法比较。结果显示,报告菌紫色杆菌CV026和根癌农杆菌A136检测发现嗜水气单胞菌和2株铜绿假单胞菌是AHLs产生菌。建立的LC-MS/MS能完全分离和检测11种AHLs,并发现假单胞菌和嗜水气单胞菌产生的AHLs信号分子含量较高,其中ATCC 9027和ATCC 15692产生3-oxo-C12-HSL,嗜水气单胞菌A2产生C4-HSL,而腐败希瓦氏菌XH4分泌的C4-HSL信号分子含量较低,沙门氏菌ATCC 14028-3、大肠杆菌O157:H7和3种弧菌的AHLs很低。随着细菌浓度的增加,铜绿假单胞菌ATCC 9027产生的AHLs含量逐步增加,在12 h达到最高。研究表明,LC-MS/MS可用于多种AHLs信号分子的定性和定量检测,具有灵敏性与准确性更高的优点。在11株水产品源致病菌和腐败菌中,假单胞菌和气单胞菌分泌的AHLs含量最高。To evaluate the N-acylated-L-homoserine lactones （AHLs）profiling of quorum sensing signal molecules of pathogen and spoilage bacteria from aquatic product, the technique of LC-MS/MS detecting simultaneously the AHLs was developed, compared with biosensor method. The results showed that Aeromonas hydrophila A2 and two strains of Pseudomonas aeruginosa secreted AHLs by biosensors Chromobacterium violacceum CV026 and Agrobacterium tumefaciens A136. The LC-MS/MS technique could be applied to qualitative and quantitative analysis of 11 kinds of AHLs. High levels of AHLs with 3- oxo-C12-HSL and C4-HSL being the most prominent were produced by P. aeruginosa and A. hydrophila, respectively. Low content of AHLs with C4-HSL was secreted by S. putrefaciens XH4, whereas no AHLs were detected in Salmonella ATCC 14028-3, Escherichia. coli O157：H7 and three species of Vibro. The production of AHLs of P. aeruginosa ATCC 9027 increased with the growth of bacteria, and the highest level of AHLs was observed after 12 h culture. Thus, the LC-MS/MS methodology can provide comprehensive profiles and concentration of AHLs in single sample, with high accuracy and sensitivity. Furthermore, P. aeruginosa and A. hydrophila produced the higher level of AHLs of quorum sensing signal molecules among 11 strains of pathogen and spoilage bacteria from aquatic product,which will provide the good foundation to explore the mechanism and control of these bacteria.

关 键 词：群体感应 AHLs 检测 LC-MS MS 

分 类 号：TS254.7[轻工技术与工程—水产品加工及贮藏工程]