Fernandes-Martins, Maria C.Springer, CarliColman, Daniel R.Boyd, Eric S.2024-10-292024-08Fernandes-Martins, M.C., Springer, C., Colman, D.R. & Boyd, E.S. (2024) Acquisition of elemental sulfur by sulfur-oxidising Sulfolobales. Environmental Microbiology, 26(9), e16691. Available from: https://doi.org/10.1111/1462-2920.166911462-2912https://scholarworks.montana.edu/handle/1/18900Elemental sulfur (S80)-oxidising Sulfolobales (Archaea) dominate high-temperature acidic hot springs (>80°C, pH <4). However, genomic analyses of S80-oxidising members of the Sulfolobales reveal a patchy distribution of genes encoding sulfur oxygenase reductase (SOR), an S80 disproportionating enzyme attributed to S80 oxidation. Here, we report the S80-dependent growth of two Sulfolobales strains previously isolated from acidic hot springs in Yellowstone National Park, one of which associated with bulk S80 during growth and one that did not. The genomes of each strain encoded different sulfur metabolism enzymes, with only one encoding SOR. Dialysis membrane experiments showed that direct contact is not required for S80 oxidation in the SOR-encoding strain. This is attributed to the generation of hydrogen sulfide (H2S) from S80 disproportionation that can diffuse out of the cell to solubilise bulk S80 to form soluble polysulfides (Sx2−) and/or S80 nanoparticles that readily diffuse across dialysis membranes. The Sulfolobales strain lacking SOR required direct contact to oxidise S80, which could be overcome by the addition of H2S. High concentrations of S80 inhibited the growth of both strains. These results implicate alternative strategies to acquire and metabolise sulfur in Sulfolobales and have implications for their distribution and ecology in their hot spring habitats.en-UScc-byhttps://creativecommons.org/licenses/by/4.0/Elemental sulfursulfur-oxidisingsulfolobalesgenomic analysesAcquisition of elemental sulfur by sulfur-oxidising SulfolobalesArticle10.1111/1462-2920.16691