The Properties and Longevity of Crude Urease Extract for Biocementation

Urease, an amidohydrolase, is an essential ingredient in the emerging engineering technique of biocementation. When free urease enzyme is used this carbonate precipitation process is often referred to as enzyme induced carbonate precipitation (EICP). To date, most engineering applications of EICP have used commercially available powdered urease. However, the high cost of commercially available urease is a major barrier to adoption of engineering applications of EICP in practice. The objective of this dissertation was to develop a simple and inexpensive enzyme production technique using agricultural resources. The specific objectives of this dissertation were (i) to develop a simple extraction process to obtain urease from common agricultural resources and identify a preferred agricultural resource for further study, (ii) to reduce the cost of enzyme production by eliminating the use of a buffer, centrifugation, and dehusking of the beans during the extraction process, (iii) investigate the stability of the extracted enzyme both in solution and after reduction to a powder by lyophilization (freeze-drying), and (iv) to study the kinetics of the extracted enzyme. The results presented in this dissertation confirmed that inexpensive crude extracts of urease from agricultural products, including jack beans, soybeans, and watermelon seeds, are effective at catalyzing urea hydrolysis for carbonate precipitation. Based upon unit yield, jack beans were identified as the preferred agricultural resource for urease extraction. Results also showed that the jack bean extract retained its activity even after replacing the buffer with tap water and eliminating acetone fractionation, centrifugation, and dehusking. It was also found that the lyophilized crude extract maintained its activity during storage for at least one year and more effectively than either the crude extract solution or rehydrated commercial urease. The kinetics of the extracted enzyme was studied to gain greater insight into the optimum concentration of urea in engineering applications of EICP. Results showed higher values for the half-saturation coefficient of the crude extract compared to the commercial enzymes. The results presented in this dissertation demonstrate the potential for a significant reduction in the cost of applying EICP in engineering practice by mass production of urease enzyme via a simple extraction process.