Rapid Metabolism of 1,4-Dioxane to below Health Advisory Levels by Thiamine-Amended Rhodococcus ruber Strain 219

Bioremediation is a promising treatment technology for 1,4-dioxane-contaminated groundwater. However, metabolic dioxane-degrading bacteria identified to date are limited by their slow kinetics and inability to sustain growth at low dioxane concentrations (<100 mu g/L). Furthermore, strains may underperform because of missing growth factors, such as amino acids or vitamins. In this work, we reevaluate Rhodococcus ruber strain 219 as a dioxane-degrading strain with bioaugmentation potential. We report rapid growth and metabolic dioxane degradation by R. ruber 219 when supplemented with thiamine (vitamin B1). We also discern that the strain lacks a complete de novo thiamine synthesis pathway, indicating that R. ruber 219 is a probable thiamine auxotroph. However, when supplemented with thiamine, the strain's Monod kinetics (K-s = 0.015 +/- 0.03 mu g/L) and exceedingly low S-min (0.49 +/- 1.16 mu g/L) suggest this strain can maintain growth at very low dioxane concentrations (<100 mu g/L). Accordingly, we demonstrate that thiamine-grown R. ruber 219 sustains degradation of dilute dioxane (<100 mu g/L) to below health advisory levels. This is the first study to report sustained metabolic dioxane biodegradation to below health advisory levels of 0.35 mu g/L. Overall, our findings solidify R. ruber 219 as a promising candidate for bioremediation of dioxane-contaminated groundwater.