Med Princ Pract. 2018;27(2):186-192. doi: 10.1159/000487306. Epub 2018 Jan 31.
The aim of this study was to purify and characterize bacteriocin from the soil isolate Bacillus subtilis GAS101, and to determine its antimicrobial as well as antibiofilm potential. The purified bacteriocin was further analyzed and evaluated for mammalian cell cytotoxicity and the possible mode of action.
Bacteriocin from B. subtilis GAS101 (an animal husbandry soil isolate) was partially purified and checked for antimicrobial and antibiofilm activity against gram-positive and gram-negative bacteria. The molecular weight of bacteriocin was determined using tricine SDS-PAGE gel. The stability of bacteriocin was investigated at various temperatures and pH levels, and its sensitivity towards 8 enzymes and 6 chemicals was determined. Cytotoxicity analysis was performed on a Vero cell line by a tetrazolium dye-based assay. Scanning electron microscopy (SEM) of bacteriocin-treated bacteria was carried out to determine the possible mode of action.
Bacteriocin from B. subtilis GAS101 was a potential inhibitor of both the indicator organisms (Staphylococcus epidermidis and Escherichia coli), and had a molecular weight of approximately 6.5 kDa. An in situ gel assay showed a zone of inhibition corresponding to the estimated protein band size. Bacteriocin was stable and showed antibacterial activity in broad ranges of temperature (30-121°C) and pH (2-12). It was sensitive to 4 proteolytic enzymes, which indicated its proteinaceous nature. Bacteriocin showed > 70% cell viability on the mammalian Vero cell line. SEM depicted that the bacteriocin was able to disrupt the bacterial cell membrane as its probable mode of action.
Thermostable and pH-tolerant bacteriocin from B. subtilis GAS101, of about 6.5 kDa, showed broad-spectrum antimicrobial and antibiofilm activity.
本研究旨在从土壤分离株枯草芽孢杆菌 GAS101 中纯化和鉴定细菌素，并确定其抗菌和抗生物膜活性。进一步分析和评估纯化的细菌素对哺乳动物细胞的细胞毒性和可能的作用方式。
从动物养殖土壤分离株枯草芽孢杆菌 GAS101 中部分纯化细菌素，并检查其对革兰氏阳性和革兰氏阴性细菌的抗菌和抗生物膜活性。使用 tricine SDS-PAGE 凝胶测定细菌素的分子量。研究了细菌素在不同温度和 pH 值下的稳定性，以及其对 8 种酶和 6 种化学物质的敏感性。通过基于四唑染料的测定法在 Vero 细胞系上进行细胞毒性分析。通过扫描电子显微镜（SEM）观察细菌素处理后的细菌，以确定可能的作用方式。
枯草芽孢杆菌 GAS101 的细菌素是指示菌（表皮葡萄球菌和大肠杆菌）的潜在抑制剂，分子量约为 6.5 kDa。原位凝胶试验显示出与估计的蛋白质带大小相对应的抑制带。细菌素在较宽的温度（30-121°C）和 pH（2-12）范围内稳定，具有抗菌活性。它对 4 种蛋白水解酶敏感，这表明其具有蛋白质性质。细菌素在哺乳动物 Vero 细胞系上的细胞活力>70%。SEM 显示，细菌素能够破坏细菌细胞膜，这可能是其作用方式。
枯草芽孢杆菌 GAS101 的热稳定和 pH 耐受细菌素，分子量约为 6.5 kDa，具有广谱抗菌和抗生物膜活性。
