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

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^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 -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 -lactam is discussed.

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