Zhao Wenqiang, Yang Shanshan, Huang Qiaoyun, Cai Peng
State Key Laboratory of Agricultural Microbiology, College of Resources and Environment, Huazhong Agricultural University, Wuhan 430070, China.
State Key Laboratory of Agricultural Microbiology, College of Resources and Environment, Huazhong Agricultural University, Wuhan 430070, China.
Colloids Surf B Biointerfaces. 2015 Apr 1;128:600-607. doi: 10.1016/j.colsurfb.2015.03.017. Epub 2015 Mar 13.
This study investigated the effect of loosely bound extracellular polymeric substances (LB-EPS) on the comprehensive surface properties of four bacteria (Bacillus subtilis, Streptococcus suis, Escherichia coli and Pseudomonas putida). The removal of LB-EPS from bacterial surfaces by high-speed centrifugation (12,000×g) was confirmed by SEM images. Viability tests showed that the percentages of viable cells ranged from 95.9% to 98.0%, and no significant difference was found after treatment (P>0.05). FTIR spectra revealed the presence of phosphodiester, carboxylic, phosphate, and amino functional groups on bacteria surfaces, and the removal of LB-EPS did not alter the types of cell surface functional groups. Potentiometric titration results suggested the total site concentrations on the intact bacteria were higher than those on LB-EPS free bacteria. Most of the acidity constants (pKa) were almost identical, except the increased pKa values of phosphodiester groups on LB-EPS free S. suis and E. coli surfaces. The electrophoretic mobilities and hydrodynamic diameters of the intact and LB-EPS free bacteria were statistically unchanged (P>0.05), indicating LB-EPS had no influence on the net surface charges and size distribution of bacteria. However, LB-ESP could enhance cell aggregation processes. The four LB-EPS free bacteria all exhibited fewer hydrophobicity values (26.1-65.0%) as compared to the intact cells (47.4-69.3%), suggesting the removal of uncharged nonpolar compounds (e.g., carbohydrates) in LB-EPS. These findings improve our understanding of the changes in cell surface characterizations induced by LB-EPS, and have important implications for assessing the role of LB-EPS in bacterial adhesion and transport behaviors.
本研究调查了松散结合的胞外聚合物(LB-EPS)对四种细菌(枯草芽孢杆菌、猪链球菌、大肠杆菌和恶臭假单胞菌)综合表面性质的影响。通过扫描电子显微镜(SEM)图像证实了通过高速离心(12,000×g)从细菌表面去除LB-EPS。活力测试表明,活细胞百分比在95.9%至98.0%之间,处理后未发现显著差异(P>0.05)。傅里叶变换红外光谱(FTIR)显示细菌表面存在磷酸二酯、羧基、磷酸根和氨基官能团,去除LB-EPS并未改变细胞表面官能团的类型。电位滴定结果表明,完整细菌上的总位点浓度高于无LB-EPS细菌上的总位点浓度。除了无LB-EPS的猪链球菌和大肠杆菌表面磷酸二酯基团的pKa值增加外,大多数酸度常数(pKa)几乎相同。完整细菌和无LB-EPS细菌的电泳迁移率和流体动力学直径在统计学上没有变化(P>0.05),表明LB-EPS对细菌的净表面电荷和大小分布没有影响。然而,LB-ESP可以增强细胞聚集过程。与完整细胞(47.4-69.3%)相比,四种无LB-EPS细菌的疏水性值均较低(26.1-65.0%),这表明去除了LB-EPS中不带电荷的非极性化合物(如碳水化合物)。这些发现增进了我们对LB-EPS诱导的细胞表面特征变化的理解,对评估LB-EPS在细菌粘附和运输行为中的作用具有重要意义。