Integrated Bioprocess and Response Surface Methodology-Based Design for Hydraulic Conductivity Reduction Using Sporosarcina pasteurii

This study examines key bioprocess parameters influencing the reduction in hydraulic conductivity in porous media via Microbially-Induced Calcite Precipitation (MICP), highlighting its relevance to environmental engineering applications such as bio-barriers and landfill liners. Sporosarcina pasteurii was utilized as the ureolytic bacterium to induce calcium carbonate precipitation under controlled laboratory conditions. Experimental variables included bacterial cell density (OD600), diameter of glass beads, concentrations of precipitation solution, bentonite, and yeast extract. A total of 42 experimental runs were conducted based on a custom design in Design-Expert software. Hydraulic conductivity was selected as the response variable to evaluate treatment performance. Response surface methodology (RSM) was applied to develop a second-order polynomial model, with statistical analyses indicating a strong model fit (R2 = 0.948, adjusted R2 = 0.929, predicted R2 = 0.868). ANOVA confirmed the significance of the main effects and interactions, particularly those involving glass bead diameter and OD600. Among the tested factors, the precipitation solution exhibited the strongest individual effect, while bentonite and yeast extract demonstrated supportive roles. Optimization revealed that a balanced combination of microbial density and chemical inputs minimized hydraulic conductivity to 0.0399 cm/s (≈95% reduction), with an overall desirability score of 1.000. Laboratory-scale experiments demonstrated field-scale applicability, underscoring the potential of biotechnological soil treatment and empirical modeling for developing sustainable low-permeability barriers.