Scholarly Work - Center for Biofilm Engineering
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Item Temperature‐dependent inactivation and catalysis rates of plant‐based ureases for engineered biomineralization(Wiley, 2020-11) Feder, Marnie J.; Akyel, Arda; Morasko, Vincent J.; Gerlach, Robin; Phillips, Adrienne J.Engineered (bio)mineralization uses the enzyme urease to catalyze the hydrolysis of urea to promote carbonate mineral precipitation. The current study investigates the influence of temperature on ureolysis rate and degree of inactivation of plant-sourced ureases over a range of environmentally relevant temperatures. Batch experiments at 30◦C demonstrated that jack bean meal (JBM) has a 1.7 to 56 times higher activity (844 μmol urea hydrolyzed g−1 JBM min−1) than the other tested plant-sourced ureases (soybean, pigeon pea and cottonseed). Hence, ureolysis and enzyme inactivation rates were evaluated for JBM at temperatures between 20◦C and 80◦C. A combined first-order urea hydrolysis and first-order enzyme inactivation model described the inactivation of urease over the investigated range of temperatures. The temperature-dependent rate coefficients (kurea) increased with temperature and ranged from 0.0018 at 20◦C to 0.0249 L g−1 JBM min−1 at 80◦C; JBM urease became ≥50% inactivated in as little as 5.2 minutes at 80◦C and in as long as 2238 minutes at 50◦C. The combined urea hydrolysis kinetics and enzyme inactivation model provides a mathematical relationship useful for the design of biomineralization technologies and can be incorporated into reactive transport modelsItem Biomineralization of Plastic Waste to Improve the Strength of Plastic-Reinforced Cement Mortar(2021-04) Kane, Seth; Thane, Abby; Espinal, Michael; Lunday, Kendra; Armagan, Hakan; Phillips, Adrienne J.; Heveran, Chelsea M.; Ryan, Cecily A.The development of methods to reuse large volumes of plastic waste is essential to curb the environmental impact of plastic pollution. Plastic-reinforced cementitious materials (PRCs), such as plastic-reinforced mortar (PRM), may be potential avenues to productively use large quantities of low-value plastic waste. However, poor bonding between the plastic and cement matrix reduces the strength of PRCs, limiting its viable applications. In this study, calcium carbonate biomineralization techniques were applied to coat plastic waste and improved the compressive strength of PRM. Two biomineralization treatments were examined: enzymatically induced calcium carbonate precipitation (EICP) and microbially induced calcium carbonate precipitation (MICP). MICP treatment of polyethylene terephthalate (PET) resulted in PRMs with compressive strengths similar to that of plastic-free mortar and higher than the compressive strengths of PRMs with untreated or EICP-treated PET. Based on the results of this study, MICP was used to treat hard-to-recycle types 3–7 plastic waste. No plastics investigated in this study inhibited the MICP process. PRM samples with 5% MICP-treated polyvinyl chloride (PVC) and mixed type 3–7 plastic had compressive strengths similar to plastic-free mortar. These results indicate that MICP treatment can improve PRM strength and that MICP-treated PRM shows promise as a method to reuse plastic waste.Item Kinetics of Calcite Precipitation by Ureolytic Bacteria under Aerobic and Anaerobic Conditions(2019-05) Mitchell, Andrew C.; Espinosa-Ortiz, Erika J.; Parks, Stacy L.; Phillips, Adrienne J.; Cunningham, Alfred B.; Gerlach, RobinThe kinetics of urea hydrolysis (ureolysis) and induced calcium carbonate (CaCO3) precipitation for engineering use in the subsurface was investigated under aerobic conditions using Sporosarcina pasteurii (ATCC strain 11859) as well as Bacillus sphaericus strains 21776 and 21787. All bacterial strains showed ureolytic activity inducing CaCO3 precipitation aerobically. Rate constants not normalized to biomass demonstrated slightly higher-rate coefficients for both ureolysis (kurea) and CaCO3 precipitation (kprecip) for B. sphaericus 21776 (kurea=0.10±0.03 h−1, kprecip=0.60±0.34 h−1) compared to S. pasteurii (kurea=0.07±0.02 h−1, kprecip=0.25±0.02 h−1), though these differences were not statistically significantly different. B. sphaericus 21787 showed little ureolytic activity but was still capable of inducing some CaCO3 precipitation. Cell growth appeared to be inhibited during the period of CaCO3 precipitation. Transmission electron microscopy (TEM) images suggest this is due to the encasement of cells and was reflected in lower kurea values observed in the presence of dissolved Ca. However, biomass regrowth could be observed after CaCO3 precipitation ceased, which suggests that ureolysis-induced CaCO3 precipitation is not necessarily lethal for the entire population. The kinetics of ureolysis and CaCO3 precipitation with S. pasteurii was further analyzed under anaerobic conditions. Rate coefficients obtained in anaerobic environments were comparable to those under aerobic conditions; however, no cell growth was observed under anaerobic conditions with NO−3, SO2−4 or Fe3+ as potential terminal electron acceptors. These data suggest that the initial rates of ureolysis and ureolysis-induced CaCO3 precipitation are not significantly affected by the absence of oxygen but that long-term ureolytic activity might require the addition of suitable electron acceptors. Variations in the ureolytic capabilities and associated rates of CaCO3 precipitation between strains must be fully considered in subsurface engineering strategies that utilize microbial amendments.Item Detecting Microbially Induced Calcite Precipitation in a Model Well-Bore Using Downhole Low-Field NMR(2017-02) Kirkland, Catherine M.; Zanetti, Sam; Grunewald, Elliot; Walsh, David O.; Codd, Sarah L.; Phillips, Adrienne J.Microbially induced calcite precipitation (MICP) has been widely researched recently due to its relevance for subsurface engineering applications including sealing leakage pathways and permeability modification. These applications of MICP are inherently difficult to monitor nondestructively in time and space. Nuclear magnetic resonance (NMR) can characterize the pore size distributions, porosity, and permeability of subsurface formations. This investigation used a low-field NMR well-logging probe to monitor MICP in a sand-filled bioreactor, measuring NMR signal amplitude and T2 relaxation over an 8 day experimental period. Following inoculation with the ureolytic bacteria, Sporosarcina pasteurii, and pulsed injections of urea and calcium substrate, the NMR measured water content in the reactor decreased to 76% of its initial value. T2 relaxation distributions bifurcated from a single mode centered about approximately 650 ms into a fast decaying population (T2 less than 10 ms) and a larger population with T2 greater than 1000 ms. The combination of changes in pore volume and surface minerology accounts for the changes in the T2 distributions. Destructive sampling confirmed final porosity was approximately 88% of the original value. These results indicate the low-field NMR well-logging probe is sensitive to the physical and chemical changes caused by MICP in a laboratory bioreactor.Item Reducing the risk of well bore leakage using engineered biomineralization barriers(2011-04) Cunningham, Alfred B.; Gerlach, Robin; Spangler, Lee H.; Mitchell, Andrew C.; Park, Saehan; Phillips, Adrienne J.If CO2 is injected in deep geological formations it is important that the receiving formation hassufficient porosity and permeability for storage and transmission and be overlain by a suitable low-permeability cap rock formation. When the resulting CO2 plume encounters a well bore, leakage may occur through various pathways in the “disturbed zone†surrounding the well casing. Gasda et al.[9], propose a method for determining effective well bore permeability from a field pressure test. If permeability results from such tests prove unacceptably large, strategies for in situ mitigation of potential leakage pathways become important. To be effective, leakage mitigation methods must block leakage pathways on timescales longer than the plume will be mobile, be able to be delivered without causing well screen plugging, and be resistant to supercritical CO2 (ScCO2) challenges. Traditional mitigation uses cement, a viscous fluid that requires a large enough aperture for delivery and that also must bond to the surrounding surfaces in order to be effective. Technologies that can be delivered via low viscosity fluids and that can effectively plug small aperture pathways, or even the porous rock surrounding the well could have significant advantages for some leakage scenarios.We propose a microbially mediated method for plugging preferential leakage pathways and/or porous media, thereby lowering the risk of unwanted upward migration of CO2, similar to thatdiscussed by Mitchell et al.[12].We examine the concept of using engineered microbial biofilms which are capable of precipitating crystalline calcium carbonate using the process of ureolysis. The resulting combination of biofilm plus mineral deposits, if targeted near points of CO2 injection, may result in the long-term sealing of preferential leakage pathways. Successful development of these biologically-based concepts could result in a CO2 leakage mitigation technology which can be applied either before CO2 injection or as a remedial measure. Results from laboratory column studies are presented which illustrate how biomineralization deposits can be developed along packed sand columns at length scales of 2.54 cm and 61 cm. Strategies for controlling mineral deposition of uniform thickness along the axis of flow are also discussed.