光动力技术对副溶血性弧菌的灭活作用  

Inactivation effect of photodynamic technology on Vibrio parahaemolyticus

作　　者：王晓迪 郑双芝 庞一 朱军莉[1] 陆海霞[1] WANG Xiaodi;ZHENG Shuangzhi;PANG Yi;ZHU Junli;LU Haixia(School of Food Science and Biotechnology,Zhejiang Gongshang University,Hangzhou 310018,China)

机构地区：[1]浙江工商大学食品与生物工程学院,浙江杭州310018

出　　处：《食品与发酵工业》2024年第3期119-125,共7页Food and Fermentation Industries

基　　金：浙江省基础公益研究计划项目(LGN22C200025)。

摘　　要：该文探究了姜黄素介导的光动力技术(photodynamic technology,PDT)协同柠檬酸处理对副溶血性弧菌的灭活效果和机理。通过平板菌落计数法测定光动力技术对副溶血性弧菌的灭活效果、扫描电镜观察细菌形态变化,酶标仪检测PDT处理后菌体活性氧(reactive oxygen species,ROS)水平以及过氧化氢酶(catalase,CAT)、超氧化物歧化酶(superoxide dismutase,SOD)、腺苷三磷酸酶(adenosine triphosphatase,ATPase)、碱性磷酸酶(alkaline phosphatase,AKP)的活性变化,琼脂糖凝胶电泳分析基因损伤程度。结果表明,PDT杀菌效果随姜黄素浓度增加、光照时间延长和柠檬酸(0.5 mg/mL)的添加而显著增强(在最高浓度姜黄素条件下P<0.05)。当姜黄素浓度为2μmol/L,柠檬酸质量浓度为0.5 mg/mL,光照2 min后,PBS中的副溶血性弧菌菌落总数从7.43 lg CFU/mL降至0。光敏剂姜黄素被蓝光激活后通过电子转移和能量转移产生具有强氧化性的ROS,菌体内ROS水平不断升高,胞内抗氧化酶(CAT、SOD)活性下降,ATPase活性下降,上清液中AKP活性上升,细菌发生严重形变甚至趋向于扁平,DNA条带变暗甚至消失,最终导致细胞死亡。综上,姜黄素介导的PDT对副溶血性弧菌有较强的杀灭作用,添加柠檬酸显著加强杀菌效果,为PDT在水产食品安全防控领域的推广应用提供了参考。In this paper,the inactivation effect and mechanism of curcumin-mediated photodynamic technology(PDT)combined with citric acid treatment on Vibrio parahaemolyticus were studied.The inactivation effect of PDT on V.parahaemolyticus was determined by flat colony counting method,and the morphological changes of bacteria were observed by scanning electron microscope.The levels of reactive oxygen species(ROS)and the activities of catalase(CAT),superoxide dismutase(SOD),adenosine triphosphatase(ATPase),alkaline phosphatase(AKP)was detected by microplate reader after PDT.The degree of gene damage was analyzed by agarose gel electrophoresis.The results showed that the germicidal effect of PDT was significantly enhanced with the increase of curcumin concentration,light time and the addition of 0.5 mg/mL citric acid(at the highest concentration of curcumin conditions P<0.05).When the concentration of curcumin was 2μmol/L,the concentration of citric acid was 0.5 mg/mL and light exposure time was 2 min,the total colony number of V.parahemolyticus in PBS decreased from 7.43 lg CFU/mL to 0 CFU/mL.As a photosensitizer,curcumin was activated by blue light to produce strong oxidizing ROS through electron and energy transfer.With the increasing level of ROS in the bacteria,the activities of CAT,SOD,and ATPase decreased,the activity of AKP in the supernatant increased,the bacteria seriously deformed and even tended to be flat,and the band of genomic DNA became darken or even disappear,eventually leading to cell death.In conclusion,curcumin-mediated PDT can effectively inactivate V.parahemolyticus,and the addition of citric acid can significantly enhance the germicidal efficacy,which provided a reference for the popularization and application of PDT in the field of aquatic food safety prevention and control.

关 键 词：光动力杀菌技术 姜黄素 柠檬酸 副溶血性弧菌 灭活作用 

分 类 号：TS254.4[轻工技术与工程—水产品加工及贮藏工程]