Cell Wall Composition of Bacillus subtilis Changes as a Function of pH and Zn2+ Exposure: Insights from Cryo-XPS Measurements

Bacteria play an important role in the biogeochemical cycling of metals in the environment. Consequently, there is an interest to understand how the bacterial surfaces interact with metals in solution and how this affects the bacterial surface. In this work we have used a surface-sensitive analysis technique, cryogenic X-ray photoelectron spectroscopy (cryo-XPS), to monitor the surface of Bacillus subtilis cells as a function of pH and Zn2+ content in saline solution. The objective of the study was twofold: (1) to investigate the agreement between two data treatment methods for XPS, as well as investigate to what extent sample pretreatment may influence XPS data of bacterial samples, and (2) to characterize how the surface chemistry of bacterial cells is influenced by different external conditions. (1) It was found that the two data treatment methods gave rise to comparable results. However, identical samples analyzed fast-frozen or dry exhibited larger differences in surface chemistry, indicating that sample pretreatment can to large extents influence the obtained surface composition of bacterial samples. (2) The bacterial cell wall (in fast-frozen samples) undergoes dramatic compositional changes with pH and with Zn2+ exposure. The compositional changes are interpreted as an adaptive metal resistance response changing the biochemical composition of the bacterial cell wall. These results have implications for how adsorption processes at the surface of bacterial cells are analyzed, understood, modeled, and predicted.