蜡样芽孢杆菌YF-2对荧光假单胞菌群体感应的淬灭活性  

The Quorum Sensing's Quenching Activity of Bacillus cereus YF-2 to Pseudomonas fluorescens

作　　者：王扬蕊 温馨 白凤翎 檀茜倩 吕欣然 励建荣 董浩 孟玉琼 马睿[4] 王树林[4] 应米燕 杨旭 Wang Yangrui;Wen Xin;Bai Fengling;Tan Xiqian;LüXinran;Li Jianrong;Dong Hao;Meng Yuqiong;Ma Rui;Wang Shulin;Ying Miyan;Yang Xu(National&Local Joint Engineering Research Center of Storage,Processing and Safety Control Technology for Fresh Agricultural and Aquatic Products,Food Safety Key Lab of Liaoning Province,School of Food Science and Engineering,Bohai University,Jinzhou 121013,Liaoning;Collaborative Innovation Center of Seafood Deep Processing,Dalian Polytechnic University,Dalian 116034,Liaoning;Shandong Meijia Group Co.Ltd.,Rizhao 276800,Shandong;Qinghai University,Xining 810016;Minze LongyangXia Aquaculture Co.,Ltd.,Hainan Tibetan Autonomous Prefecture 811800,Qinghai)

机构地区：[1]渤海大学食品科学与工程学院、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、辽宁省食品安全重点实验室,辽宁锦州121013 [2]大连工业大学、海洋食品精深加工关键技术省部共建协同创新中心,辽宁大连116034 [3]山东美佳集团有限公司,山东日照276800 [4]青海大学,西宁810016 [5]民泽龙羊峡生态水殖有限公司,青海海南藏族自治州811800

出　　处：《中国食品学报》2024年第8期187-196,共10页Journal of Chinese Institute Of Food Science and Technology

基　　金：“十三五”国家重点研发计划重点专项(2019YFD0901702)。

摘　　要：荧光假单胞菌是生鲜水产品中常见的优势腐败菌,其致腐能力受N-酰基高丝氨酸内酯(AHLs)类群体感应系统调控。以群体感应淬灭活性为导向,利用热灭活和酸敏感试验分析菌株YF-2分泌的淬灭活性物质类型。采用结晶紫染色法和激光共聚焦显微镜技术探究菌株YF-2对荧光假单胞菌生物膜的抑制和清除作用。通过生理生化和16S rDNA对菌株YF-2进行鉴定。结果表明:菌株YF-2对荧光假单胞菌AHLs具有较强的淬灭作用,降解率为100%,且淬灭活性物质在细胞提取物(CCE)中,通过热灭活及酸敏感试验初步判断淬灭活性物质为酰基转移酶或氧化还原酶。蛋白质量浓度为20,40μg/mL和80μg/mL的CCE对C4-HSL、C6-HSL及C8-HSL的降解率在52.8%~100%范围,对荧光假单胞菌生物膜的抑制率和清除率分别在25.84%~56.63%和26.12%~59.35%范围。此外,激光共聚焦图像表明CCE使荧光假单胞菌生物膜厚度减少,生物膜结构被破坏。同时,菌株YF-2被鉴定为蜡样芽孢杆菌。本研究为应用芽孢杆菌控制水产品腐败菌荧光假单胞菌提供一定的理论依据。Pseudomonas fluorescens is a dominant spoilage bacterium commonly found in fresh aquatic products,and its spoilage ability is regulated by a quorum-sensing system of N-homoserine lactones(AHLs).Based on the quorum-sensing quenching activity,the type of quenching activity substance from strain YF-2 was analyzed using heat inactivation and acid sensitivity tests.The results showed that strain YF-2 had a significant quenching effect on P.fluorescens AHLs,with a degradation rate of 100%.Its quenching active substance existed in the crude cell extract(CCE),which was preliminarily identified as acyltransferase or oxidoreductase by heat inactivation and acid sensitivity assays.The degradation of C4-HSL,C6-HSL and C8-HSL treated by the CCE at protein concentrations of 20,40μg/mL and 80μg/mL ranged from 52.8%to 100%,and the inhibition and clearance rate of P.fluorescens biofilms ranged from 25.84%to 56.63%and 26.12%to 59.35%,respectively.In addition,the confocal laser scanning microscopy showed that the CCE reduced the thickness of P.fluorescens biofilm and disrupted the biofilm structure.Meanwhile,Bacillus cereus was recognized as the source of strain YF-2.The application of B.cereus to control the spoilage bacterium P.fluorescens in aquatic goods will have a theoretical foundation thanks to this study.

关 键 词：群体感应 蜡样芽孢杆菌 荧光假单胞菌 群体感应淬灭酶 

分 类 号：TS201.3[轻工技术与工程—食品科学]