Chair of Environmental Chemistry and Bioanalytics, Faculty of Chemistry, Nicolaus Copernicus University, Gagarina 7, 87-100 Toruń, Poland.
Colloids Surf B Biointerfaces. 2013 Apr 1;104:122-7. doi: 10.1016/j.colsurfb.2012.11.018. Epub 2012 Nov 30.
Potentiometric titration and zeta potential measurements are crucial techniques for the characterization of the surface properties of bacterial cells. In this study, we investigated the effects of two commonly used electrolytes, NaNO(3) and NaClO(4), on the viability and acid-base properties of Gram-positive bacteria Bacillus subtilis. B. subtilis are non-pathogenic bacteria which are often used to model the surface properties of pathogenic microorganisms of the same genus, including Bacillus anthracis and Bacillus cereus. The survival rates of bacterial cells treated with NaNO(3) were significantly higher in comparison with microorganisms treated with NaClO(4) (5.2-6.8 and 4.1-4.7 log(10) cfu - colony-forming units, respectively). A decrease in the ionic strength (0.1 M, 0.01 M and 0.005 M) of both electrolytes increased viable bacterial cell counts in NaNO(3) treatments and decreased viable bacterial counts in NaClO(4) treatments. Potentiometric titration revealed three dominant types of cell wall functional groups: the carboxyl group (pK(a) values of 4.58-4.89), the amino group (pK(a) values of 9.62-9.89) and the phosphate group (pK(a) values of 7.12-7.49). An increase in the ionic strength of electrolytes led to a decrease in total site concentrations and a drop in buffering capacity at the examined pH values. Based on zeta potential values, measured as a function of pH and ionic strength, the isoelectric point of B. subtilis was determined at pH 2.2 for 0.005 M and 0.01 M NaNO(3). Zeta potential increased with a rise in pH, and it decreased with an increase in ionic strength.
电位滴定和动电电位测量是研究细菌细胞表面特性的关键技术。本研究考察了两种常用电解质(NaNO3和 NaClO4)对革兰氏阳性细菌枯草芽孢杆菌生存力和酸碱性质的影响。枯草芽孢杆菌是非致病性细菌,常用于模拟炭疽杆菌和蜡样芽孢杆菌等同属病原菌的表面特性。与用 NaClO4处理的微生物相比,用 NaNO3处理的细菌细胞存活率明显更高(分别为 5.2-6.8 和 4.1-4.7log10cfu-菌落形成单位)。两种电解质离子强度(0.1 M、0.01 M 和 0.005 M)降低均使 NaNO3处理中的活菌计数增加,而使 NaClO4处理中的活菌计数减少。电位滴定表明,细胞壁存在三种主要功能基团:羧基(pKa值为 4.58-4.89)、氨基(pKa值为 9.62-9.89)和磷酸基团(pKa值为 7.12-7.49)。电解质离子强度增加会导致总配体浓度降低,在考察的 pH 值下缓冲能力下降。根据作为 pH 和离子强度函数测量的动电电位值,确定枯草芽孢杆菌的等电点在 0.005 M 和 0.01 M NaNO3时为 pH 2.2。动电电位随 pH 值升高而增加,随离子强度增加而降低。