松油烯-4-醇对荧光假单胞菌抑菌能力及作用机理  

Antibacterial Activity and Mechanism of Terpinene-4-ol against Pseudomonas fluorescens

作　　者：耿一鸣 李婷婷 励建荣 谢晶[3] 林洪[4] 王轰 申照华 郭晓华 劳敏军 周小敏 于建洋 GENG Yiming;LI Tingting;LI Jianrong;XIE Jing;LIN Hong;WANG Hong;SHEN Zhaohua;GUO Xiaohua;LAO Minjun;ZHOU Xiaomin;YU Jianyang(National&Local Joint Engineering Research Center of Storage,Processing and Safety Control Technology for Fresh Agriculturaland Aquatic Products,College of Food Science and Engineering,Bohai University,Jinzhou 121013,China;College of Life Science,Dalian Minzu University,Dalian 116600,China;College of Food Science&Technology,Shanghai OceanUniversity,Shanghai 201306,China;College of Food Science and Engineering,Ocean University of China,Qingdao 266100,China;Penglai Jinglu Fishery Co.,Ltd.,Yantai 265600,China;Shandong Meijia Group Co.,Ltd.,Rizhao 276800,China;Zhejiang Xingye Industrial Group Co.,Ltd.,Zhoushan 316120,China;Rongcheng Taixiang Food Co.,Ltd.,Key Laboratory ofFrozen Prepared Marine Food Processing,Ministry of Agriculture and Rural Affairs,Weihai 264309,China)

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,辽宁锦州121013 [2]大连民族大学生命科学学院,辽宁大连116600 [3]上海海洋大学食品学院,上海201306 [4]中国海洋大学食品科学与工程学院,山东青岛266100 [5]蓬莱京鲁渔业有限公司,山东烟台265600 [6]山东美佳集团有限公司,山东日照276800 [7]浙江兴业集团有限公司,浙江舟山316120 [8]泰祥集团荣城泰祥食品股份有限公司,农业农村部冷冻调理海洋食品加工重点实验室,山东威海264309

出　　处：《食品科学》2022年第1期30-36,共7页Food Science

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

摘　　要：本实验通过测定最小抑菌剂量(minimum inhibitory concentration,MIC)及生长曲线,评价松油烯-4-醇对荧光假单胞菌的抑菌能力;随后通过碱性磷酸酶活力、电导率测定,二乙酸荧光素染色实验分析以及扫描电子显微镜观察评价松油烯-4-醇对细胞壁及细胞膜通透性的影响;最后通过测定DNA、总蛋白与胞内钠、钾离子三磷酸腺苷(adenosine triphosphate,ATP)酶活力,来进一步探究松油烯-4-醇对细胞内大分子及细胞代谢的影响。结果表明:松油烯-4-醇对荧光假单胞菌的MIC为2μL/mL;松油烯-4-醇可有效破坏荧光假单胞菌的细胞壁,破坏程度随剂量的升高而增强;松油烯-4-醇会使细胞膜的通透性增强,导致细胞内离子的外渗;扫描电子显微镜观察结果表明松油烯-4-醇能对细胞膜产生不可逆的破坏;凝胶阻滞电泳分析结果表明,松油烯-4-醇会使大分子的DNA泄露;通过测定总蛋白含量与Na+、K+-ATP酶活力可知,松油烯-4-醇能降低细胞内蛋白质的含量并能阻滞蛋白质的表达与ATP酶的合成,造成细胞凋亡。综上,松油烯-4-醇有望成为新型水产品保鲜剂,本实验可为拓展天然防腐剂在水产品的应用提供理论依据。The antibacterial activity of terpinene-4-ol against Pseudomonas fluorescens was investigated by measuring the minimum inhibitory concentration(MIC)and growth curve.Its effect on cell wall and cell membrane permeability were evaluated by determination of alkaline phosphatase activity and electrical conductivity,fluorescein diacetate(FDA)staining and scanning electron microscope(SEM)observation,and its effect on intracellular macromolecules and cellular metabolism was further explored by determining DNA,total protein and Na+,K+-ATPase activity.The results showed that the MIC was determined to be 2μL/mL.Terpinen-4-ol could effectively destroy the cell wall of P.fluorescens in a dose-dependent manner,and enhance cell membrane permeability,leading to the leakage of intracellular ions.The results of SEM showed that terpinen-4-ol could damage the cell membrane irreversibly.Gel retardation electrophoresis analysis revealed that terpinene-4-ol could cause leakage of macromolecular DNA.Additionally,terpinen-4-ol reduced intracellular protein content and blocked the expression of protein and the synthesis of ATPase,thus resulting in cell apoptosis.Therefore,terpinen-4-ol has the potential to be used as a new preservative for aquatic products.

关 键 词：松油烯-4-醇 荧光假单胞菌 抑菌活性 抑菌机理 

分 类 号：TS201.3[轻工技术与工程—食品科学]