Microbially induced CaCO3 precipitation (MICP) is considered as an alternative green technology for cement self-healing and a basis for the development of new biomaterials. However, some issues about the role of bacteria in the induction of biogenic CaCO3 crystal nucleation, growth and aggregation are still debatable. Our aims were to screen for ureolytic calcifying microorganisms and analyze their MICP abilities during their growth in urea-supplemented and urea-deficient media. Nine candidates showed a high level of urease specific activity, and a sharp increase in the urea-containing medium pH resulted in efficient CaCO3 biomineralization. In the urea-deficient medium, all ureolytic bacteria also induced CaCO3 precipitation although at lower pH values. Five strains (B. licheniformis DSMZ 8782, B. cereus 4b, S. epidermidis 4a, M. luteus BS52, M. luteus 6) were found to completely repair micro-cracks in the cement samples. Detailed studies of the most promising strain B. licheniformis DSMZ 8782 revealed a slower rate of the polymorph transformation in the urea-deficient medium than in urea-containing one. We suppose that a ureolytic microorganism retains its ability to induce CaCO3 biomineralization regardless the origin of carbonate ions in a cell environment by switching between mechanisms of urea-degradation and metabolism of calcium organic salts.
Kombucha tea was made by the fermentation of SCOBY culture of green tea broth with the addition of Fucus vesiculosus algae extract, Cetraria islandica lichen extract and their mixture. Kombucha was also made without the herbal supplements as a control. After 11 days of fermentation, in addition to the yeast Brettanomyces bruxellensis and the bacteria Komagataeibacter rhaeticus and Komagataeibacter hansenii contained in all of the samples, the yeast Zygosaccharomyces bailii and bacteria Komagataeibacter cocois were detected in the samples with the herbal extracts. In all of the kombucha with herbal additives, the total fraction of yeast was decreased as compared to the control. The total content of polyphenols and the antioxidant activity of the beverages with and without the addition of herbal extracts were comparable. The kombucha made with the algae extract showed an increased content of sucrose and organic acids, while the fructose and glucose content in the samples with algae and the mixture of extracts were lower than in the other samples. The samples with the algae extract had the highest organoleptic indicators “aroma”, “clarity” and “acidity”, while the control samples had slightly higher indicators of “taste” and “aftertaste”. The results of this study indicate the potential of algae and lichens as functional supplements for obtaining non-alcoholic fermented beverages with additional nutraceutical value.
Microbial-induced calcite precipitation (MICP) is an innovative technology in civil engineering. However, the high cost of components and the fragility of the treated soil limit its wide use. One of the possible solutions is organic waste incorporation at different stages of the technology. In the present study, we consider the use of spent brewer’s yeast (BSY) to produce bacterial inoculates and wastepaper, flax shives and sawdust as reinforcing additives into the soil. We showed that the replacement of expensive components of LB medium by BSY extract increased biomass growth characteristics of Bacillus subtilis K51, B. cereus 4b and Micrococcus luteus 6 strains by 1.4, 1.5 and 1.8 times, respectively, while for B. subtilis 168, they were comparable to LB medium. The urease activities of all strains were not reduced compared to the control. Among the three kinds of cellulose-containing waste, wastepaper incorporation into MICP-treated soil samples led to an increase in compressive strength by 2.1 times and precipitated calcite percentage by almost 1.5 times compared to a sample without additives. Thus, we showed the potential for soil stabilization through MICP using organic waste.
Biomineralization is a universal process that has implications in a variety of areas, from civil engineering to medicine. While crystallization of amorphous CaCO3 formed in vitro is known to precede the vaterite-calcite/aragonite pathway, this process could be significantly altered when induced by bacteria, particularly within the extracellular matrix (ECM) of microbial cells. We used a combination of SEM, SANS, SAXS, FTIR and XRD methods to investigate the structure of CaCO3 formed during biomineralization induced by planktonic Bacillus cereus. Formation of precipitates in the presence of CaCl2 and urea was observed both during bacterial growth and in the medium devoid of bacteria and ECM (cell-free system). The pathway for polymorphic transformations of CaCO3 from the amorphous phase to vaterite and further to calcite was confirmed for the bacterium-induced mineralization and did not depend on the concentration of Ca2+ and urea. The structure of CaCO3 sediments differed when formed in cell-free and bacterial systems and varied depending on time and the medium composition. The rate of precipitation was accelerated in the presence of DNA, which had little effect on the solid phase structure in the cell-free system, while strongly affecting the structure and polymorphic composition of the precipitates in bacterial culture.
Biocementation is a promising soil stabilization technology that relies on microbiologically induced calcite precipitation (MICP). The addition of wastepaper was found to enhance the mechanical strength of biocemented soil. This study examined the effects of incorporating wastepaper into biocemented soil, focusing on the use of the ureolytic bacterium Bacillus licheniformis DSMZ 8782 and the yeast-like fungus Scytalidium candidum 3C for soil stabilization. The optimal wastepaper content was determined to be 2%, as it did not disrupt the uniform distribution of CaCO3 and contributed to improved soil strength. The combination of bacteria and fungi significantly increased the unconfined compression strength of samples containing 2% wastepaper (161.1 kPa) compared to untreated soil (61 kPa) and bacteria-only treatments (66.5 kPa), showing improvements of 2.6 and 2.4 times, respectively. Furthermore, we demonstrated that adding fungal biomass without wastepaper significantly improved the compressive strength, achieving a value of 236.6 kPa—nine times higher than untreated soil (26.4 kPa) and four times higher than soil treated with bacteria alone (60.6 kPa). This study identifies the optimal wastepaper content and highlights the potential of combining fungal and bacterial biomass for biocementation in soil stabilization.