【作者】 王伟； 步雨珊； 杨慧； 易华西； 张兰威；

【Author】 Wang Wei;Bu Yushan;Yang Hui;Yi Huaxi;Zhang Lanwei;Department of Food Science and Engineering,Harbin Institute of Technology;College of Food Science and Engineering,Ocean University of China;

【机构】 哈尔滨工业大学食品科学与工程系； 中国海洋大学食品科学与工程学院；

【摘要】 荧光假单胞菌属于食源嗜冷菌,高温很难将其分泌的代谢物灭活,导致形成生物膜,带来持续污染。在前期基础上,利用乳酸菌抗菌肽Lac-B23开展荧光假单胞菌生物膜的消减研究。荧光假单胞菌生物膜经过抗菌肽处理后,采用激光共聚焦显微镜发现生物膜胞外多糖和蛋白质减少,其中胞外多糖含量减少达30%,表明生物膜骨架结构开始崩解;采用琼脂糖凝胶电泳测定eDNA含量变化,研究发现eDNA含量增加,表明荧光假单胞菌生物膜膜内菌体死亡导致eDNA含量增加;采用透射电镜观察到膜内菌体完整性被破坏,导致细菌内容物流出;采用激光共聚焦显微镜观察膜内菌体状态变化,发现膜内菌体死亡率明显增加;采用流式细胞仪检测生物膜内菌体凋亡状况,发现膜内菌体凋亡增多,完整性下降。以上研究结果为利用抗菌肽削减生物膜的研究与开发提供了理论依据。更多还原

【Abstract】 Pseudomonas fluorescens belongs to the food-borne psychrophilic bacteria and it is difficult to inactivate its metabolites at high temperature,which result in the formation of biofilm,causing continuous pollution.Based on early study,the antimicrobial peptide Lac-B23 was used to reduce the biofilm oi Pseudomonas fluorescens.After treating Pseudomonas fluorescens with antimicrobial peptides,laser confocal microscopy was used to observe the extracellular polysaccharide and protein,and the results showed that extracellular polysaccharide and protein decreased and the content of extracellular polysaccharide was decreased by 30%,indicating that the skeleton structure of biofilm decomposed.The content of eDNA was determined by agarose gel electrophoresis,and the results showed that the content of eDNA increased,indicating that Lac-B23 killed the bacteria within the biofilm,resulting in the increase of eDNA.Transmission electron microscopy revealed that the integrity of the biofilm was destroyed,leading to the outflow of bacterial content.Laser confocal microscopy was used to observe the changes of bacteria in the biofilm,and the results showed that the mortality rate of bacteria significantly increased.Flow cytometry was used to determine the apoptotic status of bacteria in the biofilm,and the results showed that the apoptosis of bacteria increased and the cell integrity decreased.The above results provide a theoretical basis for the research and development of biofilm subduction using antimicrobial peptides.更多还原