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Partial purification and characterization of a broad-spectrum bacteriocin produced by a Lactobacillus plantarum zrx03 isolated from infant's feces.
Food Science & Nutrition ( IF 3.5 ) Pub Date : 2020-03-18 , DOI: 10.1002/fsn3.1428 Shuang Lei 1 , Ruixiang Zhao 1 , Junliang Sun 1 , Junjian Ran 1 , Xiaoli Ruan 1 , Yang Zhu 2
Food Science & Nutrition ( IF 3.5 ) Pub Date : 2020-03-18 , DOI: 10.1002/fsn3.1428 Shuang Lei 1 , Ruixiang Zhao 1 , Junliang Sun 1 , Junjian Ran 1 , Xiaoli Ruan 1 , Yang Zhu 2
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Lactobacillus plantarum zrx03 was a bacteriocin‐producing strain isolated from infant's feces. The fermentation supernatant produced by this strain could strongly inhibit Escherichia coli JM109 ATCC 67387, Staphylococcus aureus ATCC 25923, and Listeria monocytogenes CICC 21633, in which the diameter of inhibition zone was 12.83 ± 0.62 mm, 15.08 ± 0.31 mm, 6.75 ± 0.20 mm, respectively, compared with lactic acid bacteria N1, N2, M13, M21, M31, and M37. According to amplification of 16S rRNA gene and identification of phylogenetic tree, this strain had a 1,450 bp sequence and 100% identity to the L. plantarum strain. Based on the influence of different protease treatments, such as pepsin, trypsin, papain, and proteinase K on the antimicrobial activity, this antimicrobial substance was considered to be a natural protein. Using bacteriocin produced by this strain as study object of this experiment, it had been extracted from ammonium sulfate precipitation and different organic solvents. The results showed that ethyl acetate was selected as the optimal solution to crude extraction of bacteriocin after comparing ammonium sulfate precipitation method and organic solvent extraction method, such as n‐butanol, n‐hexane, dichloromethane, trichloromethane, in which the diameter of the inhibition zones was above 28 mm. Results also showed the inhibition spectrum of the obtained bacteriocin had a broad spectrum of inhibition which could inhibit Gram‐positive, Gram‐negative, yeast. Especially, it could effectively inhibit S. aureus ATCC 25923, Bacillus subtilis CICC 10002, Bacillus anthracis CICC 20443, E. coli JM109 ATCC 67387, and Salmonella CMCC 541, and the zone diameter of inhibition has reached more than 28 mm. Moreover, it had a good thermal stability which antibacterial activity was retained 70.58% after treatment at 121°C for 30 min, and pH‐stability was between pH 2.0–9.0. These results suggested bacteriocin produced by L. plantarum zrx03 had potential application prospects in food preservation.
中文翻译：
从婴儿粪便中分离出的植物乳杆菌 zrx03 产生的广谱细菌素的部分纯化和表征。
植物乳杆菌zrx03 是从婴儿粪便中分离出来的一种产生细菌素的菌株。该菌株产生的发酵上清液对大肠杆菌JM109 ATCC 67387、金黄色葡萄球菌ATCC 25923、单核细胞增生李斯特氏菌CICC 21633具有较强的抑制作用，抑菌圈直径分别为12.83±0.62 mm、15.08±0.31 mm、6.75±0.20 mm。分别与乳酸菌N1、N2、M13、M21、M31和M37进行比较。经16S rRNA基因扩增和系统发育树鉴定，该菌株序列全长1,450 bp，与植物乳杆菌菌株同源性100%。根据胃蛋白酶、胰蛋白酶、木瓜蛋白酶、蛋白酶K等不同蛋白酶处理对抗菌活性的影响，认为这种抗菌物质是一种天然蛋白质。本实验以该菌株产生的细菌素为研究对象，通过硫酸铵沉淀和不同的有机溶剂进行提取。结果表明，通过硫酸铵沉淀法与正丁醇、正己烷、二氯甲烷、三氯甲烷等有机溶剂萃取法的比较，选择乙酸乙酯作为细菌素粗提的最佳溶液，其中抑制直径区域超过28毫米。结果还表明，所得细菌素的抑制谱具有广谱性，对革兰氏阳性、革兰氏阴性、酵母菌均有抑制作用。特别是能有效抑制金黄色葡萄球菌ATCC 25923、枯草芽孢杆菌CICC 10002、炭疽芽孢杆菌CICC 20443、大肠杆菌等。大肠杆菌JM109 ATCC 67387、沙门氏菌CMCC 541，抑菌圈直径达到28毫米以上。此外，它具有良好的热稳定性，在121℃处理30分钟后抗菌活性仍保留70.58%，pH稳定性在pH 2.0-9.0之间。这些结果表明植物乳杆菌zrx03产生的细菌素在食品保鲜方面具有潜在的应用前景。
更新日期：2020-03-18
中文翻译：
从婴儿粪便中分离出的植物乳杆菌 zrx03 产生的广谱细菌素的部分纯化和表征。
植物乳杆菌zrx03 是从婴儿粪便中分离出来的一种产生细菌素的菌株。该菌株产生的发酵上清液对大肠杆菌JM109 ATCC 67387、金黄色葡萄球菌ATCC 25923、单核细胞增生李斯特氏菌CICC 21633具有较强的抑制作用，抑菌圈直径分别为12.83±0.62 mm、15.08±0.31 mm、6.75±0.20 mm。分别与乳酸菌N1、N2、M13、M21、M31和M37进行比较。经16S rRNA基因扩增和系统发育树鉴定，该菌株序列全长1,450 bp，与植物乳杆菌菌株同源性100%。根据胃蛋白酶、胰蛋白酶、木瓜蛋白酶、蛋白酶K等不同蛋白酶处理对抗菌活性的影响，认为这种抗菌物质是一种天然蛋白质。本实验以该菌株产生的细菌素为研究对象，通过硫酸铵沉淀和不同的有机溶剂进行提取。结果表明，通过硫酸铵沉淀法与正丁醇、正己烷、二氯甲烷、三氯甲烷等有机溶剂萃取法的比较，选择乙酸乙酯作为细菌素粗提的最佳溶液，其中抑制直径区域超过28毫米。结果还表明，所得细菌素的抑制谱具有广谱性，对革兰氏阳性、革兰氏阴性、酵母菌均有抑制作用。特别是能有效抑制金黄色葡萄球菌ATCC 25923、枯草芽孢杆菌CICC 10002、炭疽芽孢杆菌CICC 20443、大肠杆菌等。大肠杆菌JM109 ATCC 67387、沙门氏菌CMCC 541，抑菌圈直径达到28毫米以上。此外，它具有良好的热稳定性，在121℃处理30分钟后抗菌活性仍保留70.58%，pH稳定性在pH 2.0-9.0之间。这些结果表明植物乳杆菌zrx03产生的细菌素在食品保鲜方面具有潜在的应用前景。
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