Water and Small-Molecule Permeation of Dormant Bacillus subtilis Spores
Author(s)Setlow, Barbara; Setlow, Peter; Knudsen, Scott; Cermak, Nathan; Delgado, Francisco Feijo; Manalis, Scott R; ... Show more Show less
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We use a suspended microchannel resonator to characterize the water and small-molecule permeability of Bacillus subtilis spores based on spores' buoyant mass in different solutions. Consistent with previous results, we found that the spore coat is not a significant barrier to small molecules, and the extent to which small molecules may enter the spore is size dependent. We have developed a method to directly observe the exchange kinetics of intraspore water with deuterium oxide, and we applied this method to wild-type spores and a panel of congenic mutants with deficiencies in the assembly or structure of the coat. Compared to wild-type spores, which exchange in approximately 1 s, several coat mutant spores were found to have relatively high water permeability with exchange times below the ∼200-ms temporal resolution of our assay. In addition, we found that the water permeability of the spore correlates with the ability of spores to germinate with dodecylamine and with the ability of TbCl₃ to inhibit germination with l-valine. These results suggest that the structure of the coat may be necessary for maintaining low water permeability.
DepartmentMassachusetts Institute of Technology. Computational and Systems Biology Program; Massachusetts Institute of Technology. Department of Biological Engineering; Massachusetts Institute of Technology. Department of Mechanical Engineering
Journal of Bacteriology
American Society for Microbiology
Knudsen, Scott M.; Cermak, Nathan; Feijó Delgado, Francisco; Setlow, Barbara; Setlow, Peter and Manalis, Scott R. “Water and Small-Molecule Permeation of Dormant Bacillus Subtilis Spores.” Edited by P. de Boer. Journal of Bacteriology 198, no. 1 (October 2015): 168–177.
Author's final manuscript