Department of Biochemistry and Molecular Biology, Pondicherry University, Puducherry, 605014, India.
Department of Biochemistry and Molecular Biology, Pondicherry University, Puducherry, 605014, India.
J Glob Antimicrob Resist. 2019 Dec;19:85-92. doi: 10.1016/j.jgar.2019.02.021. Epub 2019 Mar 7.
Catheter-associated urinary tract infections are one of the most common types of hospital-acquired infections that start with bacterial adhesion and lead to biofilm formation. The antagonistic activity of lactic acid bacteria against pathogenic organisms makes them important for medical applications.
This study evaluated the precise method for purification of bacteriocin from Lactobacillus plantarum subsp. argentoratensis SJ33, and its characterization and effectiveness for biofilm inhibition on urinary catheters coated with bacteriocin.
Purification of bacteriocin was carried out using various methods such as cell adsorption-desorption, gel permeation chromatography, and hydrophobic interaction chromatography. Bacteriocin preparation was analysed using reverse-phase high-performance liquid chromatography (HPLC) and further characterised by Tricine SDS-PAGE and Q-TOF ESI MS. Antibacterial activity of bacteriocin was assessed against 16 different Gram-positive and Gram-negative bacterial strains, and their effect on morphology was observed under scanning electron microscopy (SEM). Biofilm adherence and inhibition were evaluated by crystal violet assay, fluorescence microscopy and SEM.
Bacteriocin preparation exhibited broad-spectrum activity against both Gram-positive and Gram-negative bacteria, and SEM analysis revealed membrane pore formation. On treating with various enzymes, bacteriocin was found to be sensitive to proteases, which confirmed its proteinaceous nature. Bacteriocin showed its applicability at acidic pH in the urinary tract. Antibiofilm activity of bacteriocin established its significance in catheter-associated biofilm inhibition against Pseudomonas aeruginosa and Staphylococcus aureus. Molecular weight of bacteriocins, namely Bac F1 and Bac F2 as resolved by RP-HPLC, was estimated to be 4039 Da and 1609 Da, respectively.
导管相关尿路感染是医院获得性感染中最常见的类型之一，其起始于细菌黏附，并导致生物膜形成。乳酸菌对病原生物的拮抗活性使它们在医学应用中具有重要意义。
本研究评估了从植物乳杆菌亚种 SJ33 中精确纯化细菌素的方法，以及其对涂有细菌素的尿路导管生物膜抑制作用的特性和效果。
使用细胞吸附-解吸、凝胶渗透色谱和疏水相互作用色谱等多种方法进行细菌素的纯化。使用反相高效液相色谱（HPLC）分析细菌素制剂，并进一步通过 Tricine SDS-PAGE 和 Q-TOF ESI MS 进行特性分析。评估细菌素对 16 种不同革兰氏阳性和革兰氏阴性细菌菌株的抗菌活性，并通过扫描电子显微镜（SEM）观察其对形态的影响。通过结晶紫测定、荧光显微镜和 SEM 评估生物膜黏附和抑制作用。
细菌素制剂对革兰氏阳性和革兰氏阴性细菌均表现出广谱活性，SEM 分析显示其形成了膜孔。用各种酶处理后，细菌素对蛋白酶敏感，这证实了其蛋白质性质。细菌素在酸性 pH 值下于泌尿道中表现出适用性。细菌素的抗生物膜活性表明其在抑制与导管相关的生物膜方面具有应用潜力，可针对铜绿假单胞菌和金黄色葡萄球菌。通过 RP-HPLC 解析的细菌素 Bac F1 和 Bac F2 的分子量分别估计为 4039 Da 和 1609 Da。
