This Article Full Text Full Text (PDF) Alert me when this article is cited Alert me if a correction is posted Services Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Copyright Information Books from ASM Press MicrobeWorld Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Suzuki, T. Articles by Tahara, Y. Search for Related Content PubMed PubMed Citation Articles by Suzuki, T. Articles by Tahara, Y.

Characterization of the Bacillus subtilis ywtD Gene, Whose Product Is Involved in -Polyglutamic Acid Degradation

Department of Applied Biological Chemistry, Faculty of Agriculture, Shizuoka University, 836 Ohya, Shizuoka 422-8529, Japan

Received 21 October 2002/ Accepted 6 January 2003

The ywtD gene, which codes for an enzyme that degrades -polyglutamic acid (PGA), was cloned from Bacillus subtilis IFO16449. The gene is located immediately downstream of ywsC and ywtABC, a PGA operon involved in PGA biosynthesis, and it showed partial similarity to genes coding for DL-endopeptidase, a peptidoglycan-degrading enzyme. The ywtD gene, from which signal sequence is excised, was inserted into pET15b, and the recombinant plasmid was then transformed into Escherichia coli. Histidine-tagged YwtD was purified from sonicated cells of the transformant. The purified YwtD degraded PGA to yield two hydrolyzed products, a high-molecular-mass product (490 kDa with nearly 100% L-glutamic acid) and an 11-kDa product (with D-glutamic acid and L-glutamic acid in an 80:20 ratio). This finding and results of enzymatic analysis of the two products with carboxypeptidase G suggest that YwtD is a novel enzyme cleaving the -glutamyl bond only between D- and L-glutamic acids of PGA, and it may be designated -DL-glutamyl hydrolase.

This article has been cited by other articles:

• Fukushima, T., Kitajima, T., Yamaguchi, H., Ouyang, Q., Furuhata, K., Yamamoto, H., Shida, T., Sekiguchi, J. (2008). Identification and Characterization of Novel Cell Wall Hydrolase CwlT: A TWO-DOMAIN AUTOLYSIN EXHIBITING N-ACETYLMURAMIDASE AND DL-ENDOPEPTIDASE ACTIVITIES. J. Biol. Chem. 283: 11117-11125 [Abstract] [Full Text]  
• Candela, T., Mock, M., Fouet, A. (2005). CapE, a 47-Amino-Acid Peptide, Is Necessary for Bacillus anthracis Polyglutamate Capsule Synthesis. J. Bacteriol. 187: 7765-7772 [Abstract] [Full Text]  
• Kimura, K., Tran, L.-S. P., Uchida, I., Itoh, Y. (2004). Characterization of Bacillus subtilis {gamma}-glutamyltransferase and its involvement in the degradation of capsule poly-{gamma}-glutamate. Microbiology 150: 4115-4123 [Abstract] [Full Text]  
• Kimura, K., Tran, L.-S. P., Itoh, Y. (2004). Roles and regulation of the glutamate racemase isogenes, racE and yrpC, in Bacillus subtilis. Microbiology 150: 2911-2920 [Abstract] [Full Text]  
• Tjalsma, H., Antelmann, H., Jongbloed, J. D.H., Braun, P. G., Darmon, E., Dorenbos, R., Dubois, J.-Y. F., Westers, H., Zanen, G., Quax, W. J., Kuipers, O. P., Bron, S., Hecker, M., van Dijl, J. M. (2004). Proteomics of Protein Secretion by Bacillus subtilis: Separating the "Secrets" of the Secretome. Microbiol. Mol. Biol. Rev. 68: 207-233 [Abstract] [Full Text]