Inhibitory effects of 2-bromoethanesulfonate and protection by addition of coenzyme M in hydrogen-oxidizing marine enrichment cultures

Abstract

Since bromoethanesulfonate (BES) is an inhibitor of methane production (competitive with methyl-coenzyme M), cells able to accumulate large internal pools of methyl-coenzyme M via uptake of its precursor, HS-CoM, should be protected from BES by addition of HS-CoM to the growth medium. Hydrogen-oxidizing marine methanogen enrichments were prepared from anaerobic sediment samples collected at Sippewisset Salt Marsh and Oyster Bay Inlet near Woods Hole, MA. The three enrichments studied were a mixture of cell types with at least 50% of the culture comprised of methanogens. Methane production was found to be sensitive to BES with half maximal inhibition occurring at 5–20 μM BES depending on the enrichment. For each, half maximal protection against 40 μM BES occurred at a HS-CoM: BES molar ratio of 20 : 1 to 40 : 1. Since the protected enrichments exhibited normal sensitivity toward BES after removal of HS-CoM, it was concluded that methane production in the presence of both BES and HS-CoM resulted from true protection and not growth of BES-resistant mutants. These results suggest that uptake of HS-CoM may be a general property of methanogens occupying anaerobic marine sediments. It is possible that uptake of this coenzyme is an important nutritional feature of methanogens in their natural habitat.