The biosignature of sparite permits the distinction between gravitational cement and endostromatolites

Following a brief summary of some fundamentals in carbonate sedimentology (sedimentary petrography) that highlights the significance of organic matter, some examples of biocrystals in carbonate grains/particles, such as bioclasts or ooids, are provided as an introductory chapter to a discussion on gravitational cements versus endostromatolites. The gravitational cements, either marine (fibrous) or continental (dog-tooth), are made of hyaline (i.e., translucent) sparitic crystals whereas endostromatolites are made of colored sparitic crystals and/or micrite. Gravitational cements forms in the vadose zone whereas endostromatolites grow in small rock cavities in the marine phreatic zone. As such the latter can grow centripetally in all directions (not only downward).

Mechanism of Biomineralization Induced by Bacillus subtilis J2 and Characteristics of the Biominerals

Biomineralization induced by microorganisms has become a hot spot in the field of carbonate sedimentology; however, the mechanisms involved still need to be explored. In this study, the bacterium Bacillus subtilis J2 (GenBank MG575432) was used to induce the precipitation of calcium carbonate minerals at Mg/Ca molar ratios of 0, 3, 6, 9, and 12. Bacillus subtilis J2 bacteria released ammonia to increase pH, but the ammonia released only made the pH increase to 8.25. Carbonic anhydrase was also produced to catalyze the hydration of carbon dioxide, and this process released carbonate and bicarbonate ions that not only increased pH but also elevated carbonate supersaturation. The biominerals formed at a Mg/Ca molar ratio of 0 were spherulitic, elongated, dumbbell-shaped, and irregularly rhombohedral calcite; at a Mg/Ca molar ratio of 3, the biominerals were calcite and aragonite, the weight ratio of calcite decreased from 26.7% to 15.6%, and that of aragonite increased from 73.3% to 84.4% with increasing incubation time. At higher Mg/Ca molar ratios, the biominerals were aragonite, and the crystallinity and thermal stability of aragonite decreased with increasing Mg/Ca molar ratios. FTIR results showed that many organic functional groups were present on/within the biominerals, such as C–O–C, N–H, C=O, O–H, and C–H. HRTEM-SAED examination of the ultra-thin slices of B. subtilis J2 bacteria showed that nano-sized minerals with poor crystal structure had grown or been adsorbed on the EPS coating. The EPS of the B. subtilis J2 strain contained abundant glutamic acid and aspartic acid, which could be deprotonated in an alkaline condition to adsorb Ca2+ and Mg2+ ions; this made EPS act as the nucleation sites. This study may provide some references for further understanding of the mechanism of biomineralization induced by microorganisms.

The Significant Roles of Mg/Ca Ratio, Cl− and SO42− in Carbonate Mineral Precipitation by the Halophile Staphylococcus epidermis Y2

Carbonate precipitation induced by microorganisms has become a hot topic in the field of carbonate sedimentology, although the effects of magnesium on biomineral formation have rarely been studied. In experiments described here, magnesium sulfate and magnesium chloride were used to investigate the significant role played by Mg2+ on carbonate precipitation. In this study, Staphylococcus epidermidis Y2 was isolated and identified by 16S ribosomal DNA (rDNA) homology comparison and ammonia, pH, carbonic anhydrase, carbonate, and bicarbonate ions were monitored during laboratory experiments. The mineral phase, morphology, and elemental composition of precipitates were analyzed by XRD and SEM-EDS. Ultrathin slices of bacteria were analyzed by HRTEM-SAED and STEM. The results show that this bacterium releases ammonia and carbonic anhydrase to increase pH, and raise supersaturation via the large number of carbonate and bicarbonate ions that are released through carbon dioxide hydration catalyzed by carbonic anhydrase. The crystal cell density of monohydrocalcite is lower in a magnesium chloride medium, compared to one of magnesium sulfate. Crystals grow in the mode of a spiral staircase in a magnesium sulfate medium, but in a concentric circular pattern in a magnesium chloride medium. There was no obvious intracellular biomineralization taking place. The results presented here contribute to our understanding of the mechanisms of biomineralization, and to the role of Mg2+ in crystal form.

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, precipitation of carbonate minerals was induced by Bacillus licheniformis DB1-9 bacteria, (identity confirmed with its phylogenetic tree), to further explore the biomineralization mechanisms. During experiments, lasting up to 24 days with varying Mg/Ca molar ratios and regular monitoring of conditions, ammonia and carbonic anhydrase are released by the bacteria, resulting in a pH increase. Carbonic anhydrase could have promoted carbon dioxide hydration to produce bicarbonate and carbonate ions, and so promoted supersaturation to facilitate the precipitation of carbonate minerals. These include rhombohedral, dumbbell-shaped, and elongated calcite crystals; aragonite appears in the form of mineral aggregates. In addition, spheroidal and fusiform minerals are precipitated. FTIR results show there are organic functional groups, such as C–O–C and C=O, as well as the characteristic peaks of calcite and aragonite; these indicate that there is a close relationship between the bacteria and the minerals. Ultrathin slices of the bacteria analyzed by HRTEM, SAED, EDS, and STEM show that precipitate within the extracellular polymeric substances (EPS) has a poor crystal structure, and intracellular granular areas have no crystal structure. Fluorescence intensity and STEM results show that calcium ions can be transported from the outside to the inside of the cells. This study provides further insights to our understanding of biomineralization mechanisms induced by microorganisms.

The Significant Role of Different Magnesium: Carbonate Minerals Induced by Moderate Halophile Staphylococcus epidermis Y2

Carbonate precipitation induced by microorganism has become a hot spot in the field of carbonate sedimentology, while the effect of different magnesium on biominerals has rarely been studied. Therefore, magnesium sulfate and magnesium chloride were used to investigate the significant role played on carbonate minerals. In this study, Staphylococcus epidermidis Y2 was isolated and identified by 16S rDNA homology comparison. The ammonia, pH, carbonic anhydrase, carbonate and bicarbonate ions were investigated. The mineral phase, morphology and elemental composition were analyzed by XRD and SEM-EDS. The ultrathin slices of bacteria were analyzed by HRTEM-SAED and STEM. The result showed that this bacterium could release ammonia and carbonic anhydrase to increase pH, and elevate the supersaturation via a large number of carbonate and bicarbonate ions released through carbon dioxide hydration catalyzed by carbonic anhydrase. The crystal cell density of monohydrocalcite was lower in magnesium chloride medium than that in magnesium sulfate medium. The crystal grew in a mode of spiral staircas in magnesium sulfate medium, while in a concentric circular pattern in magnesium chloride medium. There was no obvious intracellular biomineralization. This study may be helpful to further understand the biomineralization mechanism, may also provide some references for the reconstruction of paleogeological environment.

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.