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How Moist Heat Kills Spores of Bacillus subtilis

Department of Molecular, Microbial and Structural Biology,¹ Center for Cell Analysis and Modeling, University of Connecticut Health Center, Farmington, Connecticut 06030,⁴ Department of Biology, University of Hartford, West Hartford, Connecticut 06117,² Department of Physics, East Carolina University, Greenville, North Carolina 27858³

Received 1 August 2007/ Accepted 11 September 2007

Populations of Bacillus subtilis spores in which 90 to 99.9% of the spores had been killed by moist heat gave only two fractions on equilibrium density gradient centrifugation: a fraction comprised of less dense spores that had lost their dipicolinic acid (DPA), undergone significant protein denaturation, and were all dead and a fraction with the same higher density as that of unheated spores. The latter fraction from heat-killed spore populations retained all of its DPA, but 98% of the spores could be dead. The dead spores that retained DPA germinated relatively normally with nutrient and nonnutrient germinants, but the outgrowth of these germinated spores was significantly compromised, perhaps because they had suffered damage to some proteins such that metabolic activity during outgrowth was greatly decreased. These results indicate that DPA release takes place well after spore killing by moist heat and that DPA release during moist-heat treatment is an all-or-nothing phenomenon; these findings also suggest that damage to one or more key spore proteins causes spore killing by moist heat.

Published ahead of print on 21 September 2007.

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