Effect of ethanol on BTEX biodegradation kinetics: aerobic continuous culture experiments

The use of ethanol as an automotive fuel oxygenate represents potential economic and air-quality benefits. However, little is known about how ethanol may affect the natural attenuation of petroleum product releases. Chemostat experiments were conducted with four pure cultures (representing archetypes of the known aerobic toluene degradation pathways) to determine how ethanol affects benzene, toluene, ethylbenzene, and xylene (BTEX) biodegradation kinetics. In all cases, the presence of ethanol decreased the metabolic flux of toluene (measured as the rate of toluene degradation per cell). This negative effect was counteracted by an ethanol-supported increase in biomass, which is conducive to faster degradation rates. When the influent total organic carbon (TOC) of the toluene-ethanol mixture was kept constant, the metabolic flux of toluene was proportional to its relative contribution to the influent TOC. This empirical relationship was used to derive a mathematical model that simulated effluent benzene concentrations as a function of the influent mixed-substrate composition, the dilution rate, and Monod kinetic coefficients. Under carbon-limiting conditions (1 mg/L influent benzene), the data and model simulations showed an increase in benzene removal efficiency when ethanol was fed at low concentrations (ca. 1 mg/L) because its positive effect on cell growth outweighed its negative effect on the metabolic flux of benzene. High ethanol concentrations, however, had a negative effect, causing oxygen limitation and increasing effluent benzene concentrations to higher levels than when benzene was fed alone. The slower BTEX degradation rates expected at sites with high ethanol concentrations (e.g., at gasohol-contaminated sites) could result in longer BTEX plumes and a greater risk of exposure.