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Journal of Bacteriology, October 1999, p. 6230-6237, Vol. 181, No. 20
0021-9193/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.

Characterization of a New Sigma-K-Dependent Peptidoglycan Hydrolase Gene That Plays a Role in Bacillus subtilis Mother Cell Lysis

Fajar Aji Nugroho,1 Hiroki Yamamoto,1 Yasuo Kobayashi,2 and Junichi Sekiguchi1,*

Department of Applied Biology, Faculty of Textile Science and Technology, Shinshu University, Ueda-shi, Nagano 386-8567,1 and Faculty of Agriculture, Tokyo University of Agriculture & Technology, Fuchu-shi, Tokyo 183-8509,2 Japan

Received 16 April 1999/Accepted 30 July 1999

Bacillus subtilis produces a 30-kDa peptidoglycan hydrolase, CwlH, during the late sporulation phase. Disruption of yqeE led to a complete loss of CwlH formation, indicating the identity of yqeE with cwlH. Northern blot analysis of cwlH revealed a 0.8-kb transcript after 6 to 7.5 h for the wild-type strain but not for the sigma F, sigma E, sigma G, and sigma K mutants. Expression of the sigma K-dependent cwlH gene depended on gerE. Primer extension analysis also suggested that cwlH is transcribed by Esigma K RNA polymerase. CwlH produced in Escherichia coli harboring a cwlH plasmid is an N-acetylmuramoyl-L-alanine amidase (EC 3.5.1.28) and exhibited an optimum pH of 7.0 and high-level binding to the B. subtilis cell wall. A cwlC cwlH double mutation led to a lack of mother cell lysis even after 7 days of incubation in DSM medium, but the single mutations led to mother cell lysis after 24 h.

* Corresponding author. Mailing address: Department of Applied Biology, Faculty of Textile Science and Technology, Shinshu University, 3-15-1 Tokida, Ueda-shi, Nagano 386-8567, Japan. Phone: 81 268 21 5344. Fax: 81 268 21 5331. E-mail: jsekigu{at}giptc.shinshu-u.ac.jp.

Journal of Bacteriology, October 1999, p. 6230-6237, Vol. 181, No. 20
0021-9193/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.


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