Centre for Modern Interdisciplinary Technologies Nicolaus Copernicus University, Wileńska 4, 87-100, Torun, Poland.
Department of Environmental Chemistry and Bioanalytics, Faculty of Chemistry, Nicolaus Copernicus University, Gagarina 7, 87-100, Torun, Poland.
Sci Rep. 2018 May 8;8(1):7261. doi: 10.1038/s41598-018-25024-4.
The effect of a different pH on Saccharomyces cerevisiae cells modified with calcium ions was investigated by the capillary zone electrophoresis technique. For the identification of the wild strain of S. cerevisiae, the ribosomal nucleic acid sequencing and internal transcribed spacer sequencing as well as spectrometric approach were applied. The potentiometric titration and Fourier transform infrared spectroscopy have shown the occurrence of active functional groups such as carboxyl, amine/hydroxyl, phosphate/hydrogen phosphate groups on the surface of native yeast cells. Moreover, the spectroscopy study in a medium infrared range was carried out to identify the functional groups of yeast cells that participate in calcium ions binding interaction. Furthermore, the microscopic and spectrometric analysis shows that the pH value of the calcium ions solution has a significant effect on the intensity yeast cells clumping. Additionally, the impact of yeast cell clumping on the electrophoretic behaviours was examined. The modification of surface functional groups by calcium ions significantly affected the efficiency of electrophoretic separation. However, these changes did not affect the accuracy of S. cerevisiae identification by MALDI equipment with BioTyper platform. These results form the analytical solution for coupling of electrophoresis and MALDI-TOF MS technique.
采用毛细管区带电泳技术研究了不同 pH 值对钙离子改性酿酒酵母细胞的影响。为了鉴定野生型酿酒酵母，应用了核糖体核酸测序、内部转录间隔区测序和光谱法。等电位滴定和傅里叶变换红外光谱法表明，天然酵母细胞表面存在活性官能团，如羧基、胺/羟基、磷酸/氢磷酸基团。此外，还进行了中红外范围的光谱研究，以鉴定参与钙离子结合相互作用的酵母细胞的官能团。此外，微观和光谱分析表明，钙离子溶液的 pH 值对酵母细胞聚集的强度有显著影响。此外，还研究了酵母细胞聚集对电泳行为的影响。钙离子对表面官能团的修饰显著影响了电泳分离的效率。然而，这些变化并没有影响 MALDI 设备（BioTyper 平台）对酿酒酵母鉴定的准确性。这些结果为电泳和 MALDI-TOF MS 技术的耦合提供了分析解决方案。
