Abstract
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.
Highlights
Capillary zone electrophoresis (CZE) is a well known and widely used technique enabling the separation, identification and electroanalysis of yeast or bacteria[1]
Capillary zone electrophoresis allows for the determination of a variety of biological systems such as bacteria, yeast and fungi
The characterization of microorganisms surface functional groups and its modification allowed us to develop a effective method for the electrofocusing of yeast cells
Summary
Capillary zone electrophoresis (CZE) is a well known and widely used technique enabling the separation, identification and electroanalysis of yeast or bacteria[1]. Mannoproteins are linked to polysaccharide non-covalently and CWPs are branched by β-1,6 or β-1,3 linkages of glucans[12] Such a construction of the yeast cell wall results in there being many functional groups on its surface such as phosphate, carboxyl and amino groups. Once the yeast cell has a complex wall structure and is capable to induce flocculation, the Saccharomyces cerevisiae can be considered a model of (bio)colloids. The aim of this study was to investigate the relationship between the heterogeneity and charge of the microorganism surface as well as behaviour of tested (bio)colloids during electrophoretic analysis. The impact of the cells surface modification by calcium ions on clumping of (bio)colloids and on the effectiveness of the electrophoretic mobility was examined. The spectrometric analysis of yeast modified by calcium ions may be a foundation for the coupling of capillary electrophoresis and MALDI-TOF MS analysis to eliminate the preconcentration problem of microbiological samples
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