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Journal of Bacteriology, November 2002, p. 6093-6099, Vol. 184, No. 22
0021-9193/02/$04.00+0     DOI: 10.1128/JB.184.22.6093-6099.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.

Effects of Multiple Deletions of Murein Hydrolases on Viability, Septum Cleavage, and Sensitivity to Large Toxic Molecules in Escherichia coli

Christoph Heidrich,{dagger} Astrid Ursinus, Jürgen Berger, Heinz Schwarz, and Joachim-Volker Höltje*

Max-Planck-Institut für Entwicklungsbiologie, Abteilung Biochemie, 72076 Tübingen, Germany

Received 30 May 2002/ Accepted 16 August 2002

The multiplicity of murein hydrolases found in most bacteria presents an obstacle to demonstrating the necessity of these potentially autolytic enzymes. Therefore, Escherichia coli mutants with deletions in multiple murein hydrolases, including lytic transglycosylases, amidases, and DD-endopeptidases, were constructed. Even a mutant from which seven different hydrolases were deleted was viable and grew at a normal rate. However, penicillin-induced lysis was retarded. Most of the mutants were affected in septum cleavage, which resulted in the formation of chains of cells. All three enzymes were shown to be capable of splitting the septum. Failure to cleave the septum resulted in an increase in outer membrane permeability, and thus the murein hydrolase mutants did not grow on MacConkey agar plates. In addition, the hydrolase mutants not only could be lysed by lysozyme in the absence of EDTA but also were sensitive to high-molecular-weight antibiotics, such as vancomycin and bacitracin, which are normally ineffective against E. coli.

* Corresponding author. Mailing address: Max-Planck-Institut für Entwicklungsbiologie, Abteilung Biochemie, Spemannstraße 35, 72076 Tübingen, Germany. Phone: 07071 601412. Fax: 07071 601447. E-mail: joachim-volker.hoeltje{at}tuebingen.mpg.de.

{dagger} Present address: Landesunfallkasse NRW, 40223 Düsseldorf, Germany.

Journal of Bacteriology, November 2002, p. 6093-6099, Vol. 184, No. 22
0021-9193/02/$04.00+0     DOI: 10.1128/JB.184.22.6093-6099.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.



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