凡纳滨对虾源不动杆菌群体感应信号分子分离鉴定及其调控  

Isolation,identification and regulation of quorum sensing signal molecules of Acinetobacter in Litopenaeus vannamei

作　　者：孙秀娇 朱素芹[1] 张彩丽[1] 曾名湧[1] 

机构地区：[1]中国海洋大学食品科学与工程学院,山东青岛266003

出　　处：《微生物学通报》2015年第2期437-443,共7页Microbiology China

基　　金：国家自然科学基金项目(No.31071613);十二五国家科技支撑计划项目(No.2012BAD28B05)

摘　　要：【目的】鉴定凡纳滨对虾源不动杆菌(Acinetobacter spp.M1)分泌的N-酰基高丝氨酸内酯(AHLs)类型,探究细菌生长阶段及环境因素对其分泌信号分子的影响。【方法】报告菌株平板法检测M1的AHLs的活性;采用报告平板与薄层层析(TLC)相结合法对M1分泌的AHLs类型进行鉴定。【结果】菌株M1分泌N-3-氧代-己酰基-高丝氨酸内酯和N-3-氧代-辛酰基-高丝氨酸内酯两种信号分子。在适宜条件下AHLs活性随着培养时间的延长先升高后降低,在对数末期(30 h)达到最大。弱酸和弱碱环境能够降低M1分泌AHLs的能力,p H 7.0是M1分泌AHLs的最适p H。较高浓度的Na Cl促进了个体M1分泌AHLs的能力,但是Na Cl浓度对M1总体分泌AHLs没有显著的影响。菌株M1分泌AHLs的最佳温度为30°C,温度过高或过低都会影响其分泌。【结论】菌株M1主要产生N-3-氧代-己酰基-高丝氨酸内酯和N-3-氧代-辛酰基-高丝氨酸内酯两种类型信号分子。M1的QS系统受菌体密度和环境因素的双重调控。[Objective] We detected and analyzed N-acyl-homoserine lactones （AHLs） of Acinetobacter spp. （M1） that was isolated from spoilage Litopenaeus vannamei. We also investigated the effects of bacteria growth stages and environmental factors on activities of signaling molecules. [Methods] The reporter strains （Chromobacterium violaceum CV026 and Agrobacterium tumefaciens A136） and TLC （Thin-Layer Chromatography） method were used to detect and analyze AHLs activities of M1. [Results] AHLs secreted by M1 were N-3-oxo-C6-HSL and N-3-x-oxo-Cs-HSL. The activities of AHLs increased first and then decreased with the prolongation of the culture time under appropriate culture conditions, and reached the maximum in late logarithmic stage. The ability of M1 producing AHLs was reduced by weak acid and alkalescent envirornment, the optimum pH of M1 secreting AHLs was pH 7.0. The concentration of NaC1 had no significant effect on the whole ability of M1 secreting AHLs, while high concentration promoted the ability of individual to product AHLs. The optimal temperature for strain M1 to produce AHLs was 30 ℃, and the relative higher or lower temperature inhibited the AHLs secretion. [Conclusion] The AHLs of strain M1 were N-3-oxo-C6-HSL and N-3-oxo-Cs-HSL. Quorum sensing systems of M1 was regulated by both cell density and environmental factors.

关 键 词：不动杆菌 群体感应 N-酰基高丝氨酸内酯 

分 类 号：Q93[生物学—微生物学]