The Extracellular and Intracellular Biomineralization Induced by Bacillus licheniformis DB1-9 at Different Mg/Ca Molar Ratios

Biomineralization has become a research hotspot and attracted widespread attention in the field of carbonate sedimentology. In this study, Bacillus licheniformis DB1-9 was used to induce the calcium carbonate precipitation at different magnesium calcium molar ratios in the laboratory to further explore the biomineralization mechanism. Phylogenetic tree shows that the bacteria belongs to Bacillus licheniformis species. The ammonia and carbonic anhydrase can be released by this bacteria, resulting in the pH increase, and the carbonic anhydrase can also promote the hydration reaction of carbon dioxide and subsequently produce the bicarbonate and carbonate ions to elevate the supersaturation of calcium carbonate in the liquid culture medium to facilitate the precipitation of carbonate minerals. The calcites have a shape of rhombohedron, dumbell, and elongation, and aragonite often appears in the form of mineral aggregates, besides that there are also the spherical and the fusiform minerals. FTIR result shows there are some organic functional groups, such as C-O-C and C=O, beside of the characteristic peaks of the calcite and the aragonite, indicating that microbial metabolism is closely related to the mineral formation. The superthin slices of the bacteria analyzed by HRTEM, SAED, EDS and STEM show that the surface and EPS can adsorb a large number of calcium ions and magnesium ions and EPS may act as the nucleation sites, what’s more, the intracellular nanometer-scale sphere areas show the amorphous structures, and the intracellular calcium ions and magnesium ions suggeste that they can be transported from the outside to inside the cell by diffusion along the concentration grade from high to low. This study may provide some references to further understand the biomineralization mechanism induced by microorganisms in the laboratory and the field, and also helps to explore the reason of the transition of calcite sea to aragonite sea in the geological history.