| Title |
Identification and Characterization of an Autolysin Gene, atlA, from Streptococcus criceti |
| Author |
Haruki Tamura*, Arisa Yamada, and Hirohisa Kato |
| Address |
Division of Bioregulatory Pharmacology, Department of Pharmacology, Iwate Medical University, 2-1-1 Nishitokuta, Yahaba-cho 028-3694, Japan |
| Bibliography |
Journal of Microbiology, 50(5),777-784, 2012,
|
| DOI |
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| Key Words |
degenerate PCR, homology modeling, site-directed mutagenesis, zymogram assay |
| Abstract |
AtlA of Streptococcus mutans is a major autolysin and belongs to glycoside hydrolase family 25 with cellosyl of Streptomyces coelicolor. The autolysin gene (atlA) encoding AtlA was identified from S. criceti. AtlA of S. criceti comprises the signal sequence in the N-terminus, the putative cell-wallbinding domain in the middle, and the catalytic domain in the C-terminus. Homology modeling analysis of the catalytic domain of AtlA showed the resemblance of the spatial arrangement of five amino acids around the predicted catalytic cavity to that of cellosyl. Recombinant AtlA and its four point mutants, D655A, D747A, W831A, and D849A, were evaluated on zymogram of S. criceti cells. Lytic activity was destroyed in the mutants D655A and D747A and diminished in the mutants W831A and D849A. These results suggest that Asp655 and Asp747 residues are critical for lytic activity and Trp831 and Asp849 residues are also associated with enzymatic activity. |
