摘要
目的:荧光假单胞菌通过群体感应调控相关腐败特性,如胞外蛋白酶、脂肪酶和生物被膜等,导致水产品品质下降并产生不良风味,且传统的抑菌方式极易使其产生耐药性,因此通过寻找群体感应抑制剂来干扰群体感应所调控的生理活动,可以最大程度地降低细菌的耐药性,并延长水产品的保质期。方法:以LuxI-LuxR的氨基酸序列构建同源蛋白模型并进行分子对接,选取得分前2%的小分子物质,根据其对CV026菌株产紫色菌素的抑制程度确定最佳小分子物质。通过研究其对荧光假单胞菌群集性、泳动性、胞外蛋白酶、胞外脂肪酶、胞外聚合物产量、生物被膜以及信号分子产量等群体感应表型的影响,综合评价其抑制效果。结果:分子对接得到异麦芽酮糖醇、大蒜素、芥酸甲酯及司班80共4种小分子物质,对CV026菌株产紫色菌素的抑制率分别为72.34%,12.52%,43.29%,39.37%。其中异麦芽酮糖醇能够降低荧光假单胞菌80%以上的运动能力,对胞外蛋白酶及脂肪酶产量的抑制率分别为87.13%和52.46%,显著降低其形成生物被膜的能力,抑制率可达76.25%,并且能够抑制荧光假单胞菌信号分子的表达水平。结论:异麦芽酮糖醇具有较好的群体感应抑制活性,为新型天然群体感应抑制剂的研发奠定基础。
Objectives:Pseudomonas fluorescens regulates various spoilage characteristics,including extracellular protease,lipase,and biofilm formation,through quorum sensing(QS).This leads to the deterioration of quality and the development of undesirable flavors in aquatic products.Moreover,conventional bacteriostatic methods are prone to the emergence of drug resistance.Hence,the search for quorum sensing inhibitors to disrupt the physiological activities controlled by QS offers a promising approach to minimize bacterial resistance and prolong the shelf life of aquatic products.Methods:The homologous protein model of LuxI-LuxR was constructed using the amino acid sequence.After quality assessment,molecular docking was performed.The top 2%scoring small molecule substances were selected for an experiment on inhibiting QS activity.The substance that showed the highest degree of inhibition on violacein production in strain CV026 was chosen as the best.The inhibitory effect was evaluated by studying its impact on various QS phenotypes of P.fluorescens,including swarming mobility,swimming motility,extracellular protease,extracellular lipase,extracellular polymer production,biofilm production,and signal molecule production.Results:Four small molecules,namely methyl erucate,allicin,span 80,and isomaltitol,were obtained through molecular docking.The inhibition rates of violacein production in the CV026 strain were found to be 72.34%,12.52%,43.29%,and 39.37%for these molecules,respectively.Notably,isomaltitol exhibited the ability to reduce the motility of P.fluorescein by more than 80%and inhibit the production of extracellular protease and lipase by 87.13%and 52.46%,respectively.Furthermore,isomaltitol significantly reduced the ability to form biofilm and inhibited the expression level of P.fluorescein signal molecules by 76.25%.Conclusions:Isomaltitol exhibits significant quorum sensing inhibition activity,thereby providing a solid basis for the advancement of novel natural quorum sensing inhibitors.
作者
张德福
刘可
张明
吕欣然
刘雪飞
柏雪
李学鹏
励建荣
ZHANG Defu;LIU Ke;ZHANG Ming;LÜXinran;LIU Xuefei;BAI Xue;LI Xuepeng;LI Jianrong(College of Food Science and Engineering,Bohai University,National&Local Joint Engineering Research Center of Storage,Processing and Safety Control Technology for Fresh Agricultural and Aquatic Products,Jinzhou 121013,Liaoning;Institute of Ocean Research,Bohai University,Jinzhou 121013,Liaoning)
出处
《中国食品学报》
北大核心
2025年第10期48-59,共12页
Journal of Chinese Institute of Food Science and Technology
基金
辽宁省教育厅基本科研项目(LJ212510167019)。
关键词
荧光假单胞菌
虚拟筛选
群体感应
抑制作用
Pseudomonas fluorescens
virtual screening
quorum sensing
inhibiting effects
