赖崇熙,刘唤明,贾瑞博,邓楚津,洪鹏志,赵飞

• 1:广东海洋大学食品科技学院/广东省水产品加工与安全重点实验室/广东省现代农业科技创新中心
• 2:广东恒兴饲料实业股份有限公司

摘要(Abstract)：

【目的】筛选海洋源产细菌素乳酸菌，探讨细菌素抑菌机制。【方法】从市售海鱼的肠道中筛选出一株对金黄色葡萄球菌（Staphylococcus aureus）和大肠埃希菌（Escherichia coli）有较强抑菌效果的菌株GOFEL226，通过16S rDNA序列鉴定乳酸菌，用乙酸乙酯提取细菌素，利用分光光度法、电导率法、扫描电镜技术探究细菌素的抑菌机制。【结果】菌株GOFEL226为戊糖乳植物杆菌（Lactiplantibacillus pentosus）,GFEB226细菌素对金黄色葡萄球菌和大肠埃希菌的最小抑菌质量浓度为0.883 mg/mL；其对指示菌的生长有明显的抑制作用，可导致细胞的蛋白质、核苷酸、带电离子泄漏到细胞外，对细胞膜具有破坏作用。【结论】GFEB226细菌素可通过破坏革兰阳性和革兰阴性菌的细胞膜，使其胞内物质泄漏到胞外，进而导致细胞的死亡。

关键词(KeyWords)： 海洋乳酸菌;戊糖乳植物杆菌;细菌素;抑菌机理;抑菌活性

Abstract:

Keywords:

基金项目(Foundation): 湛江市科技计划（2020A02006）

作者(Author): 赖崇熙,刘唤明,贾瑞博,邓楚津,洪鹏志,赵飞

参考文献(References)：

• [1]孙昊,唐海峰,刘欢,等.卤水中耐酸耐盐产细菌素乳酸菌的筛选[J].中国饲料, 2023(19):37-44.
• [2]高鹏,韩金志,陆兆新,等.广谱抗菌乳酸菌的分离鉴定及细菌素的提取和纯化[J].食品科学, 2016, 37(11):160-166.
• [3]方士元,谢晶.乳酸菌及其细菌素在海水鱼保鲜中应用的研究进展[J].微生物学杂志, 2019, 39(2):111-116.
• [4] RATHOD N B, PHADKE G G, TABANELLI G, et al.Recent advances in bio-preservatives impacts of lactic acid bacteria and their metabolites on aquatic food products[J].Food Bioscience, 2021, 44:101440.
• [5] KIRTONIA K, SALAUDDIN M, BHARADWAJ K K, et al.Bacteriocin:a new strategic antibiofilm agent in food industries[J]. Biocatalysis and Agricultural Biotechnology, 2021,36:102141.
• [6] WANG J S, ZHANG S W, OUYANG Y J, et al. Current developments of bacteriocins, screening methods and their application in aquaculture and aquatic products[J]. Biocatalysis and Agricultural Biotechnology, 2019, 22:101395.
• [7] DABA G M, ELNAHAS M O, ELKHATEEB W A. Beyond biopreservatives, bacteriocins biotechnological applications:history, current status, and promising potentials[J]. Biocatalysis and Agricultural Biotechnology, 2022, 39:102248.
• [8] SHENTU H F, YE P X, ZHOU Q Q, et al. Purification,characterization, and mode of action of Sakacin ZFM225, a novel bacteriocin from Lactobacillus sakei ZFM225[J].Biochemistry and Biophysics Reports, 2023, 35:101494.
• [9] MARTíN I, BARBOSA J, PEREIRA S I A, et al. Study of lactic acid bacteria isolated from traditional ripened foods and partial characterization of their bacteriocins[J]. LWT,2023, 173:114300.
• [10] YE P X, WANG J W, LIU M M, et al. Purification and characterization of a novel bacteriocin from Lactobacillus paracasei ZFM54[J]. LWT, 2021, 143:111125.
• [11] JIANG Y H, XIN W G, YANG L Y, et al. A novel bacteriocin against Staphylococcus aureus from Lactobacillus paracasei isolated from Yunnan traditional fermented yogurt:purification,antibacterial characterization, and antibiofilm activity[J].Journal of Dairy Science, 2022, 105(3):2094-2107.
• [12] LI Q B, CHEN Q Y, WU Y Y, et al. Purification, characterization and structural identification of a novel bacteriocin produced by marine original Enterococcus durans YQ-6,and its inhibition of Listeria monocytogenes[J]. LWT,2023, 173:114329.
• [13] YI L H, QI T, HONG Y, et al. Screening of bacteriocinproducing lactic acid bacteria in Chinese homemade pickle and dry-cured meat, and bacteriocin identification by genome sequencing[J]. LWT, 2020, 125:109177.
• [14] MERAL AKTA?H, ERDO?AN A,?ETIN B. Bacteriocin characterization of autochtonous Lactococcus lactis L54and its application as starter culture for Beyaz cheese[J].Food Bioscience, 2023, 53:102739.
• [15] MKADEM W, BELGUITH K, OUSSAIEF O, et al.Systematic approach to select lactic acid bacteria from spontaneously fermented milk able to fight Listeria monocytogenes and Staphylococcus aureus[J]. Food Bioscience,2023, 51:102275.
• [16] CHEN O, HONG Y, MA J H, et al. Screening lactic acid bacteria from pickle and cured meat as biocontrol agents of Penicillium digitatum on citrus fruit[J]. Biological Control, 2021, 158:104606.
• [17] HWANHLEM N, IVANOVA T, BISCOLA V, et al. Bacteriocin producing Enterococcus faecalis isolated from chicken gastrointestinal tract originating from Phitsanulok,Thailand:isolation, screening, safety evaluation and probiotic properties[J]. Food Control, 2017, 78:187-195.
• [18] SWETWIWATHANA A, VISESSANGUAN W. Potential of bacteriocin-producing lactic acid bacteria for safety improvements of traditional Thai fermented meat and human health[J]. Meat Science, 2015, 109:101-105.
• [19] ZHANG Y, YANG J M, LIU Y, et al. A novel bacteriocin PE-ZYB1 produced by Pediococcus pentosaceus zy-B isolated from intestine of Mimachlamys nobilis:purification,identification and its anti-listerial action[J]. LWT-Food Science&Technology, 2020, 118:108760.
• [20] GHANBARI M, JAMI M, KNEIFEL W, et al. Antimicrobial activity and partial characterization of bacteriocins produced by lactobacilli isolated from Sturgeon fish[J]. Food Control, 2013, 32(2):379-385.
• [21]陈全毅.海洋源抗菌活性乳酸菌筛选与细菌素分离及对单增李斯特菌作用效应的研究[D].湛江：广东海洋大学, 2021.
• [22]王娜,李慧,戴伶俐,等.产细菌素乳酸菌的筛选与鉴定[J].中国食品学报, 2020, 20(12):248-255.
• [23]孙庆申,王钰涵,韩德权,等.酸菜中具有抑菌活性乳酸菌的分离鉴定及其产细菌素特性[J].食品科学技术学报, 2022, 40(4):64-73.
• [24] VEIGA A, DA GRA?A T TOLEDO M, ROSSA L S, et al.Colorimetric microdilution assay:validation of a standard method for determination of MIC, IC50%, and IC90%of antimicrobial compounds[J]. Journal of Microbiological Methods, 2019, 162:50-61.
• [25]安俊莹.海洋源解淀粉芽孢杆菌细菌素CAMT2发酵优化、分离及抗菌机理研究[D].湛江:广东海洋大学, 2014.
• [26]黄晓英.传统发酵食品中具有抑菌特性乳酸菌的筛选、抑菌机理及其在泡菜发酵中的应用[D].成都:西南民族大学, 2022.
• [27]马欢欢.海鱼肠道中拮抗性乳酸菌筛选及在大菱鲆保鲜中的应用[D].锦州:渤海大学, 2017.
• [28]白凤翎,王扬蕊,刘佳伊,等.抗耐药性弗氏柠檬酸杆菌乳酸菌的筛选及抑菌作用[J].渤海大学学报(自然科学版), 2023, 44(1):1-9.
• [29]胡美丽.产细菌素乳酸菌的优选及在羊肉保鲜中的应用[D].石河子:石河子大学, 2023.
• [30]万倩.针对保加利亚乳杆菌的细菌素筛选鉴定、抑菌机理及应用研究[D].成都:西南民族大学, 2023.
• [31]王竞儒.片球菌素PA-1与乳酸协同抑制嗜水气单胞菌的作用与机理初探[D].天津:天津农学院, 2021.
• [32]陈瑞龙.植物乳杆菌细菌素高产菌株的诱变选育、抑菌机理及应用[D].南京:南京农业大学, 2018.
• [33] QIAO X X, DU R P, WANG Y, et al. Purification, characterization and mode of action of enterocin, a novel bacteriocin produced by Enterococcus faecium TJUQ1[J]. International Journal of Biological Macromolecules, 2020, 144:151-159.
• [34] XIN W G, WU G, YING J P, et al. Antibacterial activity and mechanism of action of bacteriocin LFX01 against Staphylococcus aureus and Escherichia coli and its application on pork model[J]. Meat Science, 2023, 196:109045.
• [35] ZHANG J, GU S B, ZHANG T R, et al. Characterization and antibacterial modes of action of bacteriocins from Bacillus coagulans CGMCC 9951 against Listeria monocytogenes[J]. LWT, 2022, 160:113272.
• [36] FU Y, ZHAO D B, WANG L Y, et al. A broad-spectrum novel bacteriocin produced by Lactobacillus sakei in Nanjing Steamed Roast Duck:purification, antimicrobial characteristics, and antibacterial mechanisms[J]. Food Bioscience, 2022, 50(Part A):101995.