Reductive dehalogenation and mineralization of 3-chlorobenzoate in the presence of sulfate by microorganisms from a methanogenic aquifer

Abstract

We investigated the anaerobic biodegradation of 3-chlorobenzoate (3CBz) by microorganisms from an aquifer where chloroaromatic compounds were previously found to resist decay in the presence of sulfate. After a lengthy lag period, 3CBz was degraded in the presence of sulfate and concurrently with sulfate reduction. Chlorine removal from 2,5- or 3,5-dichlorobenzoates and the transient appearance of benzoate from 3CBz confirmed that reductive dehalogenation was the initial fate process for these substrates. Sulfate did not influence 3CBz degradation rates in acclimated enrichment cultures but accelerated the development of 3CBz degradation activity in fresh transfers. Benzoate degradation was more rapid in the presence of sulfate regardless of the enrichment history. Nitrate, sulfite, and a headspace of air inhibited 3CBz dehalogenation, while thiosulfate had no effect. Mass balance determinations revealed that 71 to 107% of the theoretically expected amount of methane was produced from 3CBz and benzoate oxidation in the absence of sulfate. In parallel cultures containing 15 mM sulfate, methanogenesis was reduced to 48 to 71% of that theoretically expected, while sulfate reduction accounted for 12 to 50% of the reducing equivalents. In either the presence or absence of sulfate, steady-state dissolved hydrogen concentrations were similar to those reported for sulfate-reducing or methanogenic environments, respectively. Molybdate inhibited sulfate reduction and 3CBz dehalogenation to a similar extent but did not affect benzoate biodegradation. Sulfate-dependent 3CBz biodegradation was not observed. We conclude that reductive dehalogenation and sulfate reduction occur concurrently in these enrichments and that the sulfate-dependent stimulation in fresh transfers was likely due to the acceleration of benzoate oxidation.