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Item Material properties of ureolytically induced calcium carbonate adhesives(Montana State University - Bozeman, College of Engineering, 2023) Anjum, Sobia; Chairperson, Graduate Committee: Robin Gerlach; This is a manuscript style paper that includes co-authored chapters.Polymers used in adhesive applications are often petrochemical-based and release volatile organic compounds (VOCs) during application. These VOCs can accumulate indoors to the detriment of human health. Biopolymers potentially offer a non-toxic and sustainable alternative to synthetic polymers but generally have limited physical stability and low mechanical performance. One of the methods of improving the stability and adhesive performance of biopolymers is the addition of a mineral phase to reinforce biopolymer adhesives. In this work, biomineral-reinforced biopolymer adhesives were produced by ureolytically induced precipitation of calcium carbonate in the presence of guar gum and soy protein. The microbially and enzymatically induced ureolysis was carried out by the ureolytic bacterium, Sporosarcina pasteurii, or by jack bean urease. The resulting adhesives were referred to as ureolytically induced calcium carbonate precipitation (UICP)-reinforced adhesives and specifically microbially and enzymatically induced calcium carbonate (MICP and EICP)- reinforced adhesives. The adhesive strength of these composite adhesives was optimized by varying calcium and cell (or enzyme) concentrations. The adhesive strength of biomineral reinforced guar gum and soy protein biopolymers was up to 2.5 and 6 times higher than the adhesive strength of the biopolymers alone, respectively. The durability of the MICP-reinforced adhesives was tested after varying immersions (24 h and 7 days), relative humidities (50 and 80% RH), and temperatures (-20, 100, and 300?C). The durability of the MICP-reinforced adhesives, upon immersion, was significantly improved compared to biopolymer alone, and maintained their adhesive strength at moderate humidities and from below-freezing to room temperatures after 7- day exposures. To determine the effect of biopolymers on the nanoscale material properties of biomineral aggregates, enzymatically induced calcium carbonate precipitation was induced in the presence of a standard protein, Bovine Serum Albumin (BSA). Nanoindentation and Atomic Force Microscopy show that the moduli of the mineral precipitates were significantly lowered in the presence of BSA. Atomic force microscopy also showed that BSA introduced structural variations and moduli gradation in biominerals. These results demonstrate that the presence of a protein additive, specifically BSA, can alter the nanoscale structure and material properties of calcium carbonate precipitates. Using an organic additive to manipulate microscale material properties of biominerals offers possibilities for advanced control at the microscale and enhanced toughness at the macroscale for engineering applications such as in construction, binder, and adhesive applications.Item Improving pH and temperature stability of urease for ureolysis-induced calcium carbonate precipitation(Montana State University - Bozeman, College of Engineering, 2022) Akyel, Arda; Chairperson, Graduate Committee: Robin Gerlach and Adrienne Phillips (co-chair); This is a manuscript style paper that includes co-authored chapters.Ureolysis-induced calcium carbonate (CaCO 3) precipitation (UICP) is a promising technology that takes advantage of urea hydrolysis. During UICP, the enzyme urease hydrolyzes urea, and calcium carbonate can precipitate in the presence of calcium (Ca 2+). This process is also known as biomineralization, and urease is found in several bacterial and plant cells. Urease must be active to enable biomineralization engineering applications such as sealing leakage pathways around wells for CO 2 sequestration. However, biotechnological reactions are limited by physicochemical conditions (temperature, pH, toxic compounds, etc.), and conditions in practice can be suboptimal. Sporosarcina pasteurii and jack bean meal (JBM) ureolytic activities were investigated while simulating potential environmental stresses such as high temperature and pH conditions. Urease was extracted from bacterial cells to evaluate bacterial urease as an alternative to plantbased ureases. Ureolytic activities and thermal inactivation for both bacterial- and plant-based ureases were similar. Urease became thermally inactivated at elevated temperatures (> 50 °C), and urease activity also decreased when pH values moved away from circumneutral pH conditions, i.e., at pH values < 5 and > 9. Urease stability was improved through immobilization for temperatures up to 60 °C and pH values between 3.7 and 4.7. While suspended urease did not demonstrate any residual activity after a one-hour exposure to pH 4.1 at 60 °C, immobilized urease remained active after the exposure. The studies presented here suggest that UICP technology may be used in a broad range of applications, and urease stability can be improved. The use of bacterially derived urease could be cost-competitive. UICP technology not only has the potential to solve various engineering challenges, but it also has the potential to replace traditional cement technologies and contribute to a more sustainable future.