Abstract
Calcium ions are well-known as intracellular second messengers that also have an important extracellular structural role for bacteria. Recently, we found that denser biofilms were formed by Citrobacter werkmanii BF-6 in the presence of 400mM Ca2+ than that of 12.5mM Ca2+. Therefore, we employed two-dimensional (2-D) electrophoresis methods to investigate the proteome profiles of planktonic cells and biofilms in BF-6 under different concentrations of Ca2+. Meanwhile, BF-6 biofilm architecture was also visualized with confocal laser scanning microscopy (CLSM). The results demonstrated that BF-6 biofilms formed at the bottom of microtiter plates when grown in the presence of 400mM Ca2+. A total of 151 proteins from planktonic cells and biofilms after exposure of BF-6 cells to 12.5 and 400mM Ca2+ were successfully identified. Different gene ontology (GO) and KEGG pathways were categorized and enriched for the above proteins. Growth in the presence of 400mM Ca2+ induced more complex signal pathways in BF-6 than 12.5mM Ca2+. In addition, the biofilm architectures were also affected by Ca2+. Our results show two different modes of biofilm enhancement for C. werkmanii in the presence of excess Ca2+ and provide a preliminary expression of these differences based on proteomic assays.
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