Instituto Superior de Investigaciones Biológicas (INSIBIO, CONICET-UNT) and Instituto de Química Biológica, "Dr. Bernabé Bloj", Facultad de Bioquímica, Química y Farmacia, Universidad Nacional de Tucumán, Chacabuco 461, T4000ILI San Miguel de Tucumán, Argentina; Department of Analytical Chemistry, Nutrition and Food Science, School of Veterinary Sciences, University of Santiago de Compostela, 27002 Lugo, Spain.
Instituto Superior de Investigaciones Biológicas (INSIBIO, CONICET-UNT) and Instituto de Química Biológica, "Dr. Bernabé Bloj", Facultad de Bioquímica, Química y Farmacia, Universidad Nacional de Tucumán, Chacabuco 461, T4000ILI San Miguel de Tucumán, Argentina.
Int J Food Microbiol. 2021 Aug 16;352:109267. doi: 10.1016/j.ijfoodmicro.2021.109267. Epub 2021 Jun 1.
The use of bacteriocins is a promising alternative to improve food security through the biocontrol of food pathogens and spoilage microorganisms. Gram-negative produced microcin J25(G12Y), known as (MccJ25(G12Y)) is a variant of the well-studied and characterized antimicrobial peptide, microcin J25 (MccJ25). In the present work, we explored the activity of this microcin against Gram-negative bacteria linked to foodborne diseases. We evaluated the in vitro antimicrobial activity of MccJ25(G12Y) in solid medium against a collection of pathogenic and food-altering strains and studied its activity and stability in meat and dairy food systems. We show that MccJ25(G12Y) exhibited the same in vitro antimicrobial spectrum as its parental microcin (MccJ25) against different Gram-negative foodborne pathogens and spoilage strains. We highlight that low concentrations of MccJ25(G12Y) between 0.45 and 29.4 μM were able to inhibit a substantial number of pathogens, including Salmonella, Escherichia, Shigella and Enterobacter genus. We also demonstrate the antimicrobial effectiveness of the peptide against Escherichia coli O157:H7 NCTC 12900, Enterobacter cloacae CECT 194, and Salmonella enterica CECT 4396 in fish and beef burgers and yogurt. MccJ25(G12Y) was added or not to food matrices inoculated with the foodborne pathogens at 10 CFU/g or mL. Afterward, food products were stored at 4 °C and selective media for the specific enumeration were used to determine the antimicrobial susceptibility of each pathogen to MccJ25(G12Y). The viability of the three pathogens was significantly reduced in the different food biological environments. In yogurt, the peptide decreased E. coli numbers on day 5 by about 4 log 10 CFU/mL as compared to non-treated samples. For S. enterica and E. cloacae no viable cells were detected at the end of the treatment. Adding MccJ25(G12Y) to fish burgers decreased E. cloacae numbers during storage 2 log CFU/g on the first day, reaching a difference of about 5 log 10 CFU/g after 10 days compared to non-treated control. Finally, the peptide decreased E. coli O157:H7 numbers on the beef burgers samples during storage on day 10 by about 3 log 10 CFU/g as compared to non-treated samples. The stability analysis demonstrated that MccJ25(G12Y) is capable of remaining active in these food matrices for a considerable time during the storage at refrigeration temperatures. These results reinforce the studies on the potential applicability of this microcin as a biopreservative in the food industry.
使用细菌素是通过生物防治食品病原体和腐败微生物来提高食品安全的一种有前途的替代方法。革兰氏阴性菌产生的微菌素 J25(G12Y),称为(MccJ25(G12Y)),是一种经过充分研究和表征的抗菌肽微菌素 J25(MccJ25)的变体。在本工作中,我们探索了这种微菌素对与食源性疾病相关的革兰氏阴性细菌的活性。我们评估了 MccJ25(G12Y)在固体培养基中对一系列致病性和改变食物的菌株的体外抗菌活性,并研究了其在肉类和乳制品食品系统中的活性和稳定性。我们表明,MccJ25(G12Y)对不同的革兰氏阴性食源性病原体和腐败菌株的体外抗菌谱与亲代微菌素(MccJ25)相同。我们强调,低浓度的 MccJ25(G12Y)(0.45 至 29.4 μM)能够抑制大量病原体,包括沙门氏菌、大肠杆菌、志贺氏菌和肠杆菌属。我们还证明了该肽对大肠杆菌 O157:H7 NCTC 12900、阴沟肠杆菌 CECT 194 和肠炎沙门氏菌 CECT 4396 在鱼和牛肉汉堡和酸奶中的抗菌有效性。MccJ25(G12Y)被添加或不添加到接种了食源性病原体的食品基质中,浓度为 10 CFU/g 或 mL。之后,将食品产品储存在 4°C,并使用选择性培养基对每种病原体对 MccJ25(G12Y)的抗菌敏感性进行定量。在不同的食品生物环境中,三种病原体的存活能力均显著降低。在酸奶中,与未处理的样品相比,肽在第 5 天使大肠杆菌数量减少了约 4 个对数 10 CFU/mL。对于肠炎沙门氏菌和阴沟肠杆菌,在处理结束时未检测到存活细胞。在鱼汉堡中添加 MccJ25(G12Y)在第一天储存期间可使阴沟肠杆菌数量减少 2 个对数 CFU/g,10 天后与未处理的对照相比,数量减少了约 5 个对数 10 CFU/g。最后,肽在牛肉汉堡样品中的储存期间使大肠杆菌 O157:H7 的数量在第 10 天减少了约 3 个对数 10 CFU/g,与未处理的样品相比。稳定性分析表明,MccJ25(G12Y)在冷藏温度下的储存过程中能够在这些食品基质中保持相当长的时间的活性。这些结果加强了该微菌素作为食品工业中生物防腐剂的潜在应用的研究。