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Applied and Environmental Microbiology, June 2008, p. 3764-3773, Vol. 74, No. 12
0099-2240/08/$08.00+0     doi:10.1128/AEM.00453-08
Copyright © 2008, American Society for Microbiology. All Rights Reserved.

Insights into the Mode of Action of Chitosan as an Antibacterial Compound{triangledown} ,{dagger}

Dina Raafat,1* Kristine von Bargen,2 Albert Haas,2 and Hans-Georg Sahl1

Institute for Medical Microbiology, Immunology and Parasitology (IMMIP), Pharmaceutical Microbiology Unit, University of Bonn, D-53115 Bonn, Germany,1 Institute for Cell Biology, University of Bonn, D-53121 Bonn, Germany2

Received 25 February 2008/ Accepted 22 April 2008

Chitosan is a polysaccharide biopolymer that combines a unique set of versatile physicochemical and biological characteristics which allow for a wide range of applications. Although its antimicrobial activity is well documented, its mode of action has hitherto remained only vaguely defined. In this work we investigated the antimicrobial mode of action of chitosan using a combination of approaches, including in vitro assays, killing kinetics, cellular leakage measurements, membrane potential estimations, and electron microscopy, in addition to transcriptional response analysis. Chitosan, whose antimicrobial activity was influenced by several factors, exhibited a dose-dependent growth-inhibitory effect. A simultaneous permeabilization of the cell membrane to small cellular components, coupled to a significant membrane depolarization, was detected. A concomitant interference with cell wall biosynthesis was not observed. Chitosan treatment of Staphylococcus simulans 22 cells did not give rise to cell wall lysis; the cell membrane also remained intact. Analysis of transcriptional response data revealed that chitosan treatment leads to multiple changes in the expression profiles of Staphylococcus aureus SG511 genes involved in the regulation of stress and autolysis, as well as genes associated with energy metabolism. Finally, a possible mechanism for chitosan's activity is postulated. Although we contend that there might not be a single classical target that would explain chitosan's antimicrobial action, we speculate that binding of chitosan to teichoic acids, coupled with a potential extraction of membrane lipids (predominantly lipoteichoic acid) results in a sequence of events, ultimately leading to bacterial death.


* Corresponding author. Mailing address: Institute for Medical Microbiology, Immunology and Parasitology (IMMIP), Pharmaceutical Microbiology Unit, University of Bonn, Meckenheimer Allee 168, D-53115 Bonn, Germany. Phone: (49) 228 73 2113. Fax: (49) 228 73 5267. E-mail: draafat{at}uni-bonn.de

{triangledown} Published ahead of print on 2 May 2008.

{dagger} Supplemental material for this article may be found at http://aem.asm.org/.


Applied and Environmental Microbiology, June 2008, p. 3764-3773, Vol. 74, No. 12
0099-2240/08/$08.00+0     doi:10.1128/AEM.00453-08
Copyright © 2008, American Society for Microbiology. All Rights Reserved.







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