Purification and characterization of hemolysin from periodontopathogenic bacterium Eikenella corrodens strain 1073 |
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| Authors: | Fariha Jasin Mansur Sari Takahara Mihoko Yamamoto Masafumi Shimatani Mohammad Minnatul Karim Yuichiro Noiri |
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| Institution: | 1. Faculty of Agriculture, Department of Biological Chemistry, Yamaguchi University, Yamaguchi, Japan;2. Department of Biotechnology and Genetic Engineering, Islamic University, Kushtia, Bangladesh;3. Division of Cariology, Operative Dentistry and Endodontics, Department of Oral Health Science, Niigata University Graduate School of Medical and Dental Sciences, Niigata, Japan |
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| Abstract: | Eikenella corrodens 1073 was found to show hemolytic activity when grown on sheep blood agar. A high and dose-dependent hemolytic activity was detected in the cell envelope fraction, which was further purified by ion-exchange and gel-filtration chromatography. Consequently, a 65-kDa protein with hemolytic activity was obtained, suggesting that this protein might be a hemolysin. Its N-terminal amino acid sequence was nearly identical to that of X-prolyl aminopeptidase from E. corrodens ATCC 23834. To confirm that X-prolyl aminopeptidase functions as a hemolytic factor, we expressed the hlyA gene, encoding X-prolyl aminopeptidase, in Escherichia coli. After induction with isopropyl β-D-1-thiogalactopyranoside, a protein of about 65 kDa was purified on a Ni column, and its hemolytic activity was confirmed. Meanwhile, a strain with a disrupted hlyA gene, which was constructed by homologous recombination, did not show any hemolytic activity. These results suggested that X-prolyl aminopeptidase might function as a hemolysin in E. corrodens. |
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| Keywords: | Eikenella corrodens hemolysis purification virulence periodontal disease |
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