Enzyme-driven approaches to decontaminate bacillus cells and spores

Mundra, Ruchir Vishwaprakash
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Kane, Ravi S.
Dordick, Jonathan S.
Gross, Richard A.
Cramer, Steven M.
Koffas, Mattheos A. G.
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Chemical engineering
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Bacillus cells have an exposed germ cell wall and are vulnerable to irreversible enzymatic hydrolysis. However, under environmental stress, they form recalcitrant spores. Spores are highly resistant to a range of chemical and physical assaults and to the best of our knowledge, a biocatalytic sporicidal system has never been developed before. To that end, we have developed and characterized an environmentally benign biocatalytic treatment against bacillus spores. A multilayered, cross-linked coat envelops the germ cell wall and confers the spores with resistance to lytic enzymes. Taking cues from nature, we identified proteases which degrade the formidable coat layer. Proteinase K and subtilisin Carlsberg, for B. cereus and B. anthracis spore coats, respectively, led to a morphological change in the otherwise impregnable coat structure, increasing coat permeability towards cortex lytic enzymes such as lysozyme and SleB, thereby initiating spore germination. The germinated spores were further shown to be vulnerable to bacteriolytic enzymes resulting in effective spore killing.
August 2015
School of Engineering
Dept. of Chemical and Biological Engineering
Rensselaer Polytechnic Institute, Troy, NY
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