滤纸片法筛选抑制嗜水气单胞菌群体感应的乳酸菌及抑制作用分析  

Screening of Lactic Acid Bacteria with Inhibitory Activity Against Quorum Sensing of Aeromonas hydrophila Using Filter Paper Method and Their Inhibitory Effects

作　　者：林洋 孙梦桐 吕欣然[2] 白凤翎 励建荣 沈琳 LIN Yang;SUN Mengtong;Lü Xinran;BAI Fengling;LI Jianrong;SHEN Lin(Food Safety Key Lab of Liaoning Province,National ＆ Local Joint Engineering Research Center of Storage,Processing and Safety Control Technology for Fresh Agricultural and Aquatic Products,College of Food Science and Technology,Bohai University,Jinzhou 121013,China;College of Biology Sciences and Technology,Beijing Forestry University,Beijing 100083,China;Dalian Donglin Food Co.Ltd.,Dalian 116101,China)

机构地区：[1]渤海大学食品科学与工程学院辽宁省食品安全重点实验室生鲜农产品贮藏加工及安全控制技术国家地方联合工程研究中心,辽宁锦州121013 [2]北京林业大学生物科学与技术学院,北京100083 [3]大连东霖食品股份有限公司,辽宁大连116101

出　　处：《食品科学》2018年第14期125-131,共7页Food Science

基　　金："十二五"国家科技支撑计划项目(2015BAD17B05)

摘　　要：采用滤纸片法从传统发酵蔬菜中筛选抑制嗜水气单胞菌群体感应的乳酸菌菌株,应用96孔板法测定其对生物膜的抑制率,光学显微镜和扫描电镜观察其对嗜水气单胞菌生物膜形成的影响,同时以蛋白酶、嗜铁素、群集及泳动为指标研究其对嗜水气单胞菌毒力作用的影响。结果表明:来源于辽宁锦州酸菜的乳酸菌菌株SCT-2对嗜水气单胞菌群体感应有明显抑制作用,当其代谢产物粗提物质量浓度为8 mg/mL时,对嗜水气单胞菌生物膜抑制率为45.16%,光学显微镜下显示菌株SCT-2粗提物可有效抑制嗜水气单胞菌生物膜的形成,扫描电镜结果进一步表明菌株SCT-2粗提物不仅降低了嗜水气单胞菌生物膜的生成量,而且使其生物膜断裂。8 mg/mL的SCT-2粗提物可使嗜水气单胞菌蛋白酶和嗜铁素的分泌量分别减少27.18%和22.11%,对信号分子的降解率为32.27%,且对嗜水气单胞菌的群集和泳动现象抑制明显。经生理生化和16S rRNA鉴定菌株SCT-2为植物乳杆菌(Lactobacillus plantarum)。从传统东北酸菜中获得对嗜水气单胞菌群体感应有抑制作用的植物乳杆菌SCT-2,为开发一种抑制嗜水气单胞菌群体感应的乳酸菌生物制剂提供了理论支持。Lactic acid bacteria（LAB） with inhibitory activity against quorum sensing of Aeromonas hydrophila were screened from traditional fermented vegetables using the filter paper method. The biofilm inhibitory activity of the LAB isolates was measured by a 96-well microtiter plate reader, and their effect on the biofilm formation of A. hydrophila was observed by light microscopy and scanning electron microscopy（SEM）. The proteolytic activity, siderophore production, swarming and swimming were used as indicators to evaluate the effects of these quorum sensing inhibitors（QSI） on the virulence of A. hydrophila. In this study, a strain designated SCT-2, with inhibitory activity toward quorum sensing of A. hydrophila, was isolated from pickled cabbage from Jinzhou, Liaoning province. Crude metabolite extract from strain SCT-2 at 8 mg/mL inhibited the biofilm formation of A. hydrophila by 45.16%. Light microscopy revealed that the crude extract pronouncedly inhibited the biofilm formation of A. hydrophila. The SEM images also revealed that the crude extract not only decreased the production of biofilm of A. hydrophila, but also destroyed the biofilm. The secretion amount of proteolytic and siderophore in A. hydrophila treated with 8 mg/mL of the crude extract was decreased by 27.18% and 22.11%, respectively. In addition, the crude extraction degraded 32.27% of the N-acylhomoserine lactones（AHLs） of A. hydrophila. It also inhibited obviously the swarming and swimming of A. hydrophila. Strain SCT-2 was identified as Lactobacillus plantarum by physiological and biochemical tests and 16 S rRNA sequence analysis. This study may pave the foundation for the development LAB-based biological agent for inhibiting quorum sensing of A. hydrophila.

关 键 词：传统发酵酸菜 乳酸菌 群体感应抑制剂 嗜水气单胞菌 生物膜 

分 类 号：TS201.3[轻工技术与工程—食品科学]