Abstract
Chlorinated aliphatic hydrocarbons (CAHs), such as cis-1,2-dichloroethylene (cDCE), are prevalent in groundwater at many locations throughout the United States. When immobilized in hydrogel beads with slow-release compounds, the bacteria strain Rhodococcus rhodochrous ATCC 21198 can be used for the in situ bioremediation of cDCE. These hydrogel beads must exhibit high mechanical strength and resist degradation to extend the lifetime of slow-release compounds and bioremediation. We engineered poly(vinyl)-alcohol - alginate (PVA-AG) beads to immobilize ATCC 21198 with the slow-release compound, tetrabutoxysilane (TBOS) that produces 1-butanol as a growth substrate, for high mechanical strength. We optimized three inputs (concentration of PVA, concentration of AG, and the crosslinking time) on two responses (compressive modulus and rate of oxygen utilization) for batch incubation experiments between 1 and 30 days using a design of experiments approach. The predictive models generated from design of experiments were then tested by measuring the compressive strength, oxygen utilization, and abiotic rates of hydrolysis for a predicted optimal bead formulation. The result of this study generated a hydrogel bead with immobilized R. rhodochrous ATCC 21198 and TBOS that exhibited a high compressive modulus on day 1 and day 30, which was accurately predicted by models. These hydrogel beads exhibited low metabolic activity based on oxygen rates on day 1 and day 30 but were not accurately predicted by the models. In addition, the ratio between oxygen utilization and abiotic rates of hydrolysis were observed to be roughly half of what was expected stoichiometrically. Lastly, we demonstrated the capability to use these beads as a bioremediation technology for cDCE as we found that, for all bead formulations, cDCE was significantly reduced after 30 days. Altogether, this work demonstrates the capability to capture and enhance the material properties of the complex hydrogel beads with predictive models yet signals the need for more robust methods to understand the metabolic activity that occurs in the hydrogel beads.