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从发酵菠萝(Ananas comosus (L.) Merr.)中分离出的大肠杆菌E产生的部分纯化细菌素的特性鉴定与安全性评估

Characterization and safety evaluation of partially purified bacteriocin produced by Escherichia coli E isolated from fermented pineapple Ananas comosus (L.) Merr.

作者信息

Le Van Thi, Leelakriangsak Montira, Lee Seong Wei, Panphon Somrak, Utispan Kusumawadee, Koontongkaew Sittichai

机构信息

Biology Division, Department of Science, Faculty of Science and Technology, Prince of Songkla University, Pattani, 94000, Thailand.

Faculty of Agro Based Industry, Universiti Malaysia Kelantan Campus Jeli, Jeli, Kelantan, 17600, Malaysia.

出版信息

Braz J Microbiol. 2019 Jan;50(1):33-42. doi: 10.1007/s42770-018-0014-5. Epub 2018 Dec 4.

Abstract

Antibacterial activity of cell-free supernatant from Escherichia coli E against selected pathogenic bacteria in food and aquaculture was the highest against Edwardsiella tarda 3, a significant aquaculture pathogen. Biochemical properties of the bacteriocins were studied and bacteriocin was found to be sensitive to proteinase K, demonstrating its proteinaceous nature. In addition, pH and temperature affected bacteriocin activity and stability. The bacteriocins were partially purified by ammonium sulfate precipitation. The antibacterial activity was only detected in 20% ammonium sulfate fraction and direct detection of its activity was performed by overlaying on the indicator strains. The inhibition zone associated with the antibacterial activity was detected in the sample overlaid by E. tarda 3 and Staphylococcus aureus DMST8840 with the relative molecular mass of about 27 kDa and 10 kDa, respectively. Bacteriocin showed no cytotoxic effect on NIH-3T3 cell line; however, two virulence genes, aer and sfa, were detected in the genome of E. coli E by PCR. The characteristics of bacteriocins produced by E. coli E exhibited the antibacterial activity against both Gram-positive and Gram-negative pathogenic bacteria and the safe use determined by cytotoxicity test which may have interesting biotechnological applications.

摘要

大肠杆菌E的无细胞上清液对食品和水产养殖中选定的病原菌的抗菌活性对迟钝爱德华氏菌3最高,迟钝爱德华氏菌3是一种重要的水产养殖病原菌。研究了细菌素的生化特性,发现细菌素对蛋白酶K敏感,表明其蛋白质性质。此外,pH值和温度影响细菌素的活性和稳定性。通过硫酸铵沉淀对细菌素进行了部分纯化。抗菌活性仅在20%硫酸铵级分中检测到,并通过覆盖指示菌株直接检测其活性。在分别覆盖迟钝爱德华氏菌3和金黄色葡萄球菌DMST8840的样品中检测到与抗菌活性相关的抑菌圈,其相对分子质量分别约为27 kDa和10 kDa。细菌素对NIH-3T3细胞系无细胞毒性作用;然而,通过PCR在大肠杆菌E的基因组中检测到两个毒力基因aer和sfa。大肠杆菌E产生的细菌素的特性表现出对革兰氏阳性和革兰氏阴性病原菌的抗菌活性,并且通过细胞毒性试验确定其安全使用,这可能具有有趣的生物技术应用。

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