摘要
为探究草鱼鱼鳞抗菌肽与肉桂精油联合作用的抑菌机理,本研究采用Tricine-十二烷基硫酸钠-聚丙烯酰胺凝胶电泳法测定草鱼鱼鳞抗菌肽的分子质量,通过棋盘稀释法分析草鱼鱼鳞抗菌肽与肉桂精油联合抑菌效果;与此同时,对经复配抑菌剂处理的受试菌生长曲线、细胞膜渗透性、胞外乳酸脱氢酶活力进行测定,并利用流式细胞仪分析穿膜效率。结果表明:分子质量约为14.3 kDa的草鱼鱼鳞抗菌肽与肉桂精油联用对金黄色葡萄球菌、副溶血性弧菌、产黄青霉的抑菌效果表现为协同作用,对沙门氏菌、大肠杆菌、黑曲霉、毛霉、根霉菌、酵母菌的抑菌效果表现为相加作用,对所有受试菌的抑菌效果均无拮抗作用。通过流式细胞仪分析发现,复配抑菌剂对金黄色葡萄球菌的穿膜效率为72.76%、对产黄青霉为61.56%、对副溶血性弧菌为51.04%,均明显高于单独使用抑菌剂。复配抑菌剂的抑菌机理可能是通过影响菌体的生长周期,致使菌体对数生长期缩短以及衰亡期加快;通过改变细胞膜通透性,使得胞内电解质、乳酸脱氢酶不断溢出到胞外,细胞生长受到抑制,进而导致死亡。
In this study,tricine-sodium dodecyl sulfate-polyacrylamide gel electrophoresis(TSDS-PAGE)was used to determine the molecular mass of an antimicrobial peptide from grass carp scale.Chessboard dilution method was used to determine the combined antimicrobial effect of the peptide and cinnamon essential oil on common foodborne pathogenic bacteria,and the growth curves,cell membrane permeability,and extracellular lactate dehydrogenase activity of the tested strains when exposed to combinations of the two antimicrobials were measured.In addition,flow cytometry was used to analyze the membrane penetration efficiency.The results showed that combination of the peptide,with a molecular mass of 14.3 kDa,and cinnamon essential oil exerted a synergistic effect against Staphylococcus aureus,Vibrio parahaemolyticus and Penicillium chrysogenum and additive effect against Salmonella choleraesuis,Escherichia coli,Aspergillus niger,Mucor wutungkiao,Rhizopus and Saccharomycetes,but had no antagonistic effect on all tested bacteria.Through flow cytometry analysis,it was found that the membrane penetration efficiency of the antimicrobial combination was 72.76%for Staphylococcus aureus,61.56%for Penicillium chrysogenum,and 51.04%for Vibrio parahaemolyticus,all of which were significantly higher than those of either antimicrobial alone.The antimicrobial mechanism may be by shortening the logarithmic growth period,accelerating the occurrence of the decline period,changing the permeability of the cell membrane,and consequently leading to leakage of intracellular electrolytes and lactate dehydrogenase and subsequent cell death.
作者
王雪燕
陈瑛
张嘉敏
施永清
WANG Xueyan;CHEN Ying;ZHANG Jiamin;SHI Yongqing(School of Food Science and Biotechnology,Zhejiang Gongshang University,Hangzhou 310018,China)
出处
《食品科学》
EI
CAS
CSCD
北大核心
2020年第23期100-106,共7页
Food Science
基金
食品科学与工程浙江省重中之重一级学科资助项目(2017SIAR221)
浙江工商大学研究生科研创新基金项目(18020080066)。
关键词
抗菌肽
精油
联合抑菌活性
抑菌机理
antimicrobial peptide
essential oil
combined antimicrobial activity
antimicrobial mechanism