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Journal of Bacteriology, November 2002, p. 6007-6015, Vol. 184, No. 21
0021-9193/02/$04.00+0     DOI: 10.1128/JB.184.21.6007-6015.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.

A Polysaccharide Deacetylase Gene (pdaA) Is Required for Germination and for Production of Muramic {delta}-Lactam Residues in the Spore Cortex of Bacillus subtilis

Tatsuya Fukushima,1 Hiroki Yamamoto,1 Abdelmadjid Atrih,2 Simon J. Foster,2 and Junichi Sekiguchi1*

Department of Applied Biology, Faculty of Textile Science and Technology, Shinshu University, Ueda-shi, Nagano 386, Japan,1 Department of Molecular Biology and Biotechnology, University of Sheffield, Fifth Court, Western Bank, Sheffield S10 2TN, United Kingdom2

Received 15 April 2002/ Accepted 31 July 2002

The predicted amino acid sequence of Bacillus subtilis yfjS (renamed pdaA) exhibits high similarity to those of several polysaccharide deacetylases. ß-Galactosidase fusion experiments and results of Northern hybridization with sporulation sigma mutants indicated that the pdaA gene is transcribed by E{sigma}G RNA polymerase. pdaA-deficient spores were bright by phase-contrast microscopy, and the spores were induced to germination on the addition of L-alanine. Germination-associated spore darkening, a slow and partial decrease in absorbance, and slightly lower dipicolinic acid release compared with that by the wild-type strain were observed. In particular, the release of hexosamine-containing materials was lacking in the pdaA mutant. Muropeptide analysis indicated that the pdaA-deficient spores completely lacked muramic {delta}-lactam. A pdaA-gfp fusion protein constructed in strain 168 and pdaA-deficient strains indicated that the protein is localized in B. subtilis spores. The biosynthetic pathway of muramic {delta}-lactam is discussed.

* 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: 81-268-21-5345. E-mail: jsekigu{at}giptc.shinshu-u.ac.jp.

Journal of Bacteriology, November 2002, p. 6007-6015, Vol. 184, No. 21
0021-9193/02/$04.00+0     DOI: 10.1128/JB.184.21.6007-6015.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.



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