The use of principle component analysis and MALDI-TOF MS for the differentiation of mineral forming Virgibacillus and Bacillus species isolated from sabkhas

Occurrence of mineral forming and other bacteria in mats is well demonstrated. However, their high diversity shown by ribotyping has not been explained, although it could explain the diversity of formed minerals. Common biomarkers as well as phylogenic relationships are useful tools for clustering the isolates and the prediction of their potential role in the natural niche. In this study, a combination of MALDI-TOF MS with PCA was shown to be a powerful tool to categorize 35 mineral forming bacterial isolates isolated from Dohat Faishakh sabkha, northwest of Qatar (23 from decaying mats and 12 from living ones). The 23 strains from decaying mats belong to the Virgibacillus genus as identified by ribotyping and are shown to be highly involved in the formation of protodolomite and a diversity of minerals. They were used as internal references for the categorization of sabkha bacteria. Combination of the isolation of bacteria on selective mineral forming media, their MALDI TOF MS protein profiling and PCA analysis established their relationship in a phylloproteomic dendrogram based on protein biomarkers including m/z 4905, 3265, 5240, 6430, 7765, and 9815. PCA analysis clustered the studied isolates into 3 major clusters, showing strong correspondence to the 3 phylloproteomic groups that were established by the dendrogram. Both clustering analysis means have evidently demonstrated a relationship between known Virgibacillus strains and other related bacteria based on profiling of their synthesized proteins. Thus, larger populations of bacteria in mats can be easily screened for their potential to exhibit certain activities, which is of ecological, environmental and biotechnological significance.