食品与发酵工业 >

2024 , Vol. 50 >Issue 21: 51 - 59

DOI: https://doi.org/10.13995/j.cnki.11-1802/ts.037784

研究报告

栀子花乙酸乙酯萃取物对副溶血性弧菌抑菌活性及机理研究

  • 胡钰 ,
  • 张齐杰 ,
  • 傅纤雯 ,
  • 王艳 ,
  • 许光治 ,
  • 张有做 ,
  • 倪勤学
展开
  • 1(浙江农林大学 食品与健康学院,浙江 杭州,311300)
    2(浙江骄栀科技有限公司,浙江 杭州,311300)
第一作者:硕士研究生(倪勤学副教授为通信作者,E-mail:niqinxue@zafu.edu.cn)

收稿日期: 2023-10-28

  修回日期: 2024-01-02

  网络出版日期: 2024-11-28

基金资助

浙江省大学生科技活动计划暨新苗人才计划资助项目(2022R412A036)

收起

Bacteriostatic activity and mechanism of ethyl acetate extract from Gardenia jasminoides Ellis flower against Vibrio parahaemolyticus

  • HU Yu ,
  • ZHANG Qijie ,
  • FU Qianwen ,
  • WANG Yan ,
  • XU Guangzhi ,
  • ZHANG Youzuo ,
  • NI Qinxue
Expand
  • 1(Food and Health College, Zhejiang A&F University, Hangzhou 311300, China)
    2(Zhejiang Jiaozhi Technology Co.Ltd., Hangzhou 311300, China)

Received date: 2023-10-28

  Revised date: 2024-01-02

  Online published: 2024-11-28

Fold

摘要

为探究栀子花乙酸乙酯相(Gardenia jasminoides Eills flower- ethyl acetate extract,GFE-EA)对副溶血性弧菌(Vibrio parahemolyticus)的抑菌活性及机理,测定了GFE-EA对V.parahemolyticus的最低抑菌浓度(minimal inhibit concentration,MIC)和最低杀菌浓度(minimum bactericidal concentration,MBC)。通过对其生长曲线、细胞内容物泄露、细胞壁膜通透性、流动性和完整性、胞内酶活性,三磷酸腺苷(adenosine triphosphate,ATP)、丙二醛(malondialdehyde,MDA)和活性氧(reactive oxygen species,ROS)含量等的影响来探究其可能存在的抑菌机理。结果表明,GFE-EA对V.parahemolyticus的MIC和MBC分别达到2.5、20 mg/mL。与高于MIC浓度的GFE-EA共培养,发现细菌的生长受到明显抑制,上清液中核酸和蛋白质含量有显著的增加,扫描电镜发现菌体细胞形态变化明显,细胞膜流动性、通透性和完整性变差,V.parahemolyticus胞内抗氧化酶及ATP酶活性均呈下降趋势,MDA含量上升,ROS水平升高,并且其DNA荧光强度变弱,电泳条带弥散。说明GFE-EA能够通过延迟缩短细菌生长的对数期,破坏细菌细胞壁膜系统和细胞形态,影响胞内能量代谢,破坏菌体遗传物质等来抑制细菌的生长繁殖,甚至导致其死亡。该研究为探明GFE-EA抑制食源性病原菌的机理奠定了理论基础,表明栀子花在天然食品防腐剂领域中具有广泛的开发应用潜力。

关键词： 栀子花; 副溶血性弧菌; 抑菌机理; 扫描电子显微镜; DNA损伤

本文引用格式

胡钰 , 张齐杰 , 傅纤雯 , 王艳 , 许光治 , 张有做 , 倪勤学 . 栀子花乙酸乙酯萃取物对副溶血性弧菌抑菌活性及机理研究[J]. 食品与发酵工业, 2024 , 50(21) : 51 -59 . DOI: 10.13995/j.cnki.11-1802/ts.037784

Abstract

This study aimed to investigate the bacteriostatic activity and mechanism of GFE-EA from Gardenia jasminoides Eills flower against Vibrio parahaemolyticus.The study determined the minimum inhibitory concentration and minimum bactericidal concentration of GFE-EA against V.parahemolyticus.The possible antibacterial mechanism was explored by growth curve, cell content leakage, cell wall and membrane permeability, fluidity and integrity, intracellular enzyme activity, adenosine triphosphate, malondialdehyde, reactive oxygen species (ROS) content, and intracellular DNA structure.Results showed that the MIC and MBC reached 2.5 mg/mL and 20 mg/mL of GFE-EA against V.parahemolyticus, respectively.When co-cultured with GFE-EA of higher than MIC, it was found that the growth of bacteria was significantly inhibited and the contents of nucleic acid and protein in the supernate were significantly increased.The morphology of bacterial cells was significantly changed by scanning electron microscopy.The fluidity, permeability, and integrity of cell membranes deteriorated.The activity of antioxidant enzymes and ATPase in V.parahemolyticus decreased, MDA content increased and ROS level increased.The DNA fluorescence intensity was weakened and the bands dispersed, which could achieve the purpose of inhibiting bacterial growth and death by delaying and shortening the logarithmic growth period of bacteria, damaging bacterial cell wall membrane system and cell morphology, affecting their energy metabolism, and destroying bacterial genetic material.This study laid a theoretical foundation for exploring the mechanism of GFE-EA inhibiting food-borne pathogens, indicating that Gardenia jasminoides Ellis has a wide potential for development and application in the field of natural food preservatives.

Key words： Gardenia jasminoides Ellis; Vibrio parahaemolyticus; antibacterial mechanism; scanning electron microscope; DNA damage

参考文献

[1] NELAPATI S, NELAPATI K, CHINNAM B.Vibrio parahaemolyticus- An emerging foodborne pathogen[J].Veterinary World, 2012:48.
[2] LOPATEK M, WIECZOREK K, OSEK J.Prevalence and antimicrobial resistance of Vibrio parahaemolyticus isolated from raw shellfish in Poland[J].Journal of Food Protection, 2015, 78(5):1029-1033.
[3] ABANTO M, GAVILAN R G, BAKER-AUSTIN C, et al.Global expansion of Pacific northwest Vibrio parahaemolyticus sequence type 36[J].Emerging Infectious Diseases, 2020, 26(2):323-326.
[4] 陈江, 陈莉莉, 王绩凯, 等.浙江省食源性病例副溶血性弧菌检出率时空聚集分析[J].中国公共卫生, 2022, 38(11):1379-1381.
CHEN J, CHEN L L, WANG J K, et al.Spatio-temporal clustering of Vibrio parahaemolyticus detection rate among foodborne disease cases in Zhejiang Province, 2016—2017:A monitoring data analysis[J].Chinese Journal of Public Health, 2022, 38(11):1379-1381.
[5] YÜZBAŞıOĞLU D, ÜNAL F, KOÇ F, et al.Genotoxicity assessment of vaccine adjuvant squalene[J].Food and Chemical Toxicology, 2013, 56:240-246.
[6] 刘文伟, 沈晓盛, 刘承初.茶多酚对泥蚶中副溶血性弧菌的抑制效果[J].食品与发酵工业, 2010, 36(5):41-44.
LIU W W, SHEN X S, LIU C C.Effect of Tea Polyphenol to Vibrio parahaemolyticus in Tegillarca granosas[J].Food and Fermentation Industries, 2010, 36(5):41-44.
[7] PALAMAE S, MITTAL A, YINGKAJORN M, et al.Vibrio parahaemolyticus isolates from Asian green mussel:Molecular characteristics, virulence and their inhibition by chitooligosaccharide-tea polyphenol conjugates[J].Foods, 2022, 11(24):4048.
[8] 谢庆超, 孙晓红, 沈潇, 等.蓝莓提取物对副溶血性弧菌的抑制作用[J].天然产物研究与开发, 2012, 24(8):1094-1097;1108.
XIE Q C, SUN X H, SHEN X, et al.Bacteriostasis effects of blueberries extracts on Vibrio parahaemolyticus[J].Natural Product Research and Development, 2012, 24(8):1094-1097;1108.
[9] 卢路路, 樊怡灵, 邓珂, 等.不同品种和花期栀子花挥发性物质的主成分和聚类分析[J].核农学报, 2021, 35(7):1601-1608.
LU L L, FAN Y L, DENG K, et al.Principal component and cluster analysis of volatile components in cape jasmine flower from different cultivars at different stages of bloom[J].Journal of Nuclear Agricultural Sciences, 2021, 35(7):1601-1608.
[10] 阙珊奇, 杨良缘, 胡钰, 等.花期及提取方法对重瓣栀子花成分及活性影响[J].核农学报, 2023, 37(6):1193-1200.
QUE S Q, YANG L Y, HU Y, et al.Effects of flowering period and extraction methods on the ingredients and activity of Gardenia flower[J].Journal of Nuclear Agricultural Sciences, 2023, 37(6):1193-1200.
[11] 钟宜科, 吴迪, 花晓丹, 等.黄酮类化合物抑菌作用研究进展[J].中国食品添加剂, 2019, 30(8):166-171.
ZHONG Y K, WU D, HUA X D, et al.Progress in the study of antibacterial effect of flavonoids compounds[J].China Food Additives, 2019, 30(8):166-171.
[12] WANG L Y, GUO H Y, WANG J, et al.Effects of Herba Lophatheri extract on the physicochemical properties and biological activities of the chitosan film[J].International Journal of Biological Macromolecules, 2019, 133:51-57.
[13] TRACZEWSKI M M, AMBLER J E, SCHUCH R.Determination of MIC quality control parameters for exebacase, a novel lysin with antistaphylococcal activity[J].Journal of Clinical Microbiology, 2021, 59(7):e0311720.
[14] 高瑞芳, 王占黎, 曹星月, 等.内蒙古香青兰提取物的抑菌作用及机制研究[J].中医药学报, 2023, 51(2):37-41.
GAO R F, WANG Z L, CAO X Y, et al.Bacteriostatic effect and mechanism of Dracocephalum moldavica L.extracts in Inner Mongolia[J].Acta Chinese Medicine and Pharmacology, 2023, 51(2):37-41.
[15] SUN Y, WANG X D, ZHU J L, et al.Antibacterial activity and mechanism of lemon grass essential oil (LG-EO)on Vibrio parahaemolyticus[J].Journal of Chinese Institute of Food Science and Technology, 2023, 23(3):157-167.
[16] 周志娥, 罗秋水, 熊建华, 等.绿原酸、异绿原酸A对大肠杆菌的抑菌机制[J].食品科技, 2014, 39(3):228-232.
ZHOU Z E, LUO Q S, XIONG J H, et al.Antimicrobial mechanisms of 3-O-caffeoyl quinic acid and 3, 5-di-O-caffeoyl quinic acid against Escherichia coli[J].Food Science and Technology, 2014, 39(3):228-232.
[17] 王铭遥, 郑明静, 任中阳, 等.凡纳滨对虾抗菌肽的筛选及其抑菌活性和DNA结合机制研究[J].中国食品学报, 2024,23(7):140-151.
WANG M Y, ZHENG M J, REN Z Y, et al.Screening of antimicrobial peptides from Penaeus vannamei to investigate antimicrobial activity and DNA binding mechanism[J].Journal of Chinese Institute of Food Science and Technology, 2024,23(7):140-151.
[18] 孙畅, 李湘, 王银红, 等.山苍子精油对柑橘意大利青霉的抑菌活性及作用机制[J].食品科学, 2023, 44(11):17-25.
SUN C, LI X, WANG Y H, et al.Antibacterial activity and mechanism of Litsea cubeba essential oil against Penicillium italicum in Citrus fruits[J].Food Science, 2023, 44(11):17-25.
[19] ZHOU Y D, ZHANG Y L, ZONG H, et al.Chemical constituents, antibacterial activity and mechanism of Paeonia suffruticosa Andr.buds extract against Staphylococcus aureus and Escherichia coli O157:H7[J].Natural Product Research, 2021, 35(6):1005-1009.
[20] 王虎玄, 彭中花, 王聪, 等.香芹酚对鲁氏接合酵母的抗菌活性及其机制[J].中国食品学报, 2023, 23(2):27-39.
WANG H X, PENG Z H, WANG C, et al.The antifungal activity and mechanism of carvacrol against Zygosaccharomyces rouxii[J].Journal of Chinese Institute of Food Science and Technology, 2023, 23(2):27-39.
[21] 王奇升, 但阿康, 罗海涛, 等.石花菜醇提物提取工艺及抗氧化、抗菌活性研究[J].中国食品学报, 2023, 23(1):171-183.
WANG Q S, DAN A K, LUO H T, et al.Studies on the extraction process, antioxidant and antibacterial activity of Gelidium amansii ethanol extract[J].Journal of Chinese Institute of Food Science and Technology, 2023, 23(1):171-183.
[22] 刘雪飞, 吴思楠, 李秀霞, 等.细胞壁缺失对副溶血性弧菌生物学特性的影响[J].中国食品学报, 2021, 21(5):23-30.
LIU X F, WU S N, LI X X, et al.Effects of cell wall deletion on the biological characteristics of Vibrio parahaemolyticus[J].Journal of Chinese Institute of Food Science and Technology, 2021, 21(5):23-30.
[23] 谢丽玲, 彭齐, 蔡链纯, 等.黄芩醇提物对副溶血性弧菌抑制机制的研究[J].生物技术通报, 2015(8):159-165.
XIE L L, PENG Q, CAI L C, et al.A study on antibacterial mechanisms of Ethanol-extracts from Scutellaria baicalensis AgainstVibrio parahaemolyticus[J].Biotechnology Bulletin, 2015(8):159-165.
[24] 王晓迪, 郑双芝, 庞一, 等.光动力技术对副溶血性弧菌的灭活作用[J].食品与发酵工业, 2024, 50(3):119-125.
WANG X D, ZHENG S Z, PANG Y, et al.Inactivation effect of photodynamic technology on Vibrio parahaemolyticus[J].Food and Fermentation Industries, 2024, 50(3):119-125.
[25] PALAMAE S, MITTAL A, BUATONG J, et al.Chitooligosaccharide-catechin conjugate:Antimicrobial mechanisms toward Vibrio parahaemolyticus and its use in shucked Asian green mussel[J].Food Control, 2023, 151:109794.
Options
文章导航

/

〈 〉

技术支持：北京玛格泰克科技发展有限公司