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Antimicrobial Agents and Chemotherapy, December 2008, p. 4281-4288, Vol. 52, No. 12
0066-4804/08/$08.00+0     doi:10.1128/AAC.00625-08
Copyright © 2008, American Society for Microbiology. All Rights Reserved.

Inhibition of Bacillus anthracis Spore Outgrowth by Nisin

Ian M. Gut,¹ Angela M. Prouty,¹ Jimmy D. Ballard,⁴ Wilfred A. van der Donk,^2,3* and Steven R. Blanke^1,3*

Department of Microbiology, University of Illinois, Urbana, Illinois 61801,¹ Department of Chemistry, University of Illinois, Urbana, Illinois 61801,² Institute for Genomic Biology, University of Illinois, Urbana, Illinois 61801,³ Department of Microbiology and Immunology, The University of Oklahoma Health Sciences Center, Oklahoma City, Oklahoma 73104⁴

Received 13 May 2008/ Returned for modification 17 July 2008/ Accepted 10 September 2008

The lantibiotic nisin has previously been reported to inhibit the outgrowth of spores from several Bacillus species. However, the mode of action of nisin responsible for outgrowth inhibition is poorly understood. By using B. anthracis Sterne 7702 as a model, nisin acted against spores with a 50% inhibitory concentration (IC₅₀) and an IC₉₀ of 0.57 µM and 0.90 µM, respectively. Viable B. anthracis organisms were not recoverable from cultures containing concentrations of nisin greater than the IC₉₀. These studies demonstrated that spores lose heat resistance and become hydrated in the presence of nisin, thereby ruling out a possible mechanism of inhibition in which nisin acts to block germination initiation. Rather, germination initiation is requisite for the action of nisin. This study also revealed that nisin rapidly and irreversibly inhibits growth by preventing the establishment of oxidative metabolism and the membrane potential in germinating spores. On the other hand, nisin had no detectable effects on the typical changes associated with the dissolution of the outer spore structures (e.g., the spore coats, cortex, and exosporium). Thus, the action of nisin results in the uncoupling of two critical sequences of events necessary for the outgrowth of spores: the establishment of metabolism and the shedding of the external spore structures.

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* Corresponding author. Mailing address for Wilfred A. van der Donk: Department of Chemistry, University of Illinois, Urbana, IL 61801. Phone: (217) 244-5360. Fax: (217) 244-8533. E-mail: vddo.nk{at}illinois.edu. Mailing address for Steven R. Blanke: Department of Microbiology, University of Illinois, Urbana, IL 61801. Phone (217) 244-2412. Fax: (217) 244-6697. E-mail: sblanke{at}life.uiuc.edu

Published ahead of print on 22 September 2008.

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Antimicrobial Agents and Chemotherapy, December 2008, p. 4281-4288, Vol. 52, No. 12
0066-4804/08/$08.00+0     doi:10.1128/AAC.00625-08
Copyright © 2008, American Society for Microbiology. All Rights Reserved.

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