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Purification and biochemical characterization of bacteriolytic enzyme from Bacillus subtilis YU-1432 active against Porphyromonas gingivalis

Journal of the Korean Society for Applied Biological Chemistry volume 54, pages 600–605 (2011)Cite this article

Abstract

YU205, a bacteriolytic enzyme produced by Bacillus subtilis YU-1432 exhibited lytic activity against Porphyromonas gingivalis causing periodontal disease. Specific activity and purification yield of YU205 were increased by 522.0 times and 21.9%, respectively by 80% acetone precipitation, followed by DEAE-Sepharose and CM-Sepharose column chromatography. The molecular mass of YU205 was estimated to be 29.0 kDa by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), and N-terminal sequencing identified fifteen amino acid residues, AQSVPYGISQIKAPA. YU205 was stabilized against thermal inactivation at 1 mM of CaCl2. The essential amino acids for the lytic activity were deduced to be tyrosine, methionine, and serine. YU205 showed at least 23.4 times higher substrate specificity to P. Gingivalis than other proteolytic enzymes tested. These results suggest that the purified enzyme may have potential for the application to food or dental care products to prevent periodontal diseases.

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Authors and Affiliations

  1. Department of Bio-Food Materials, College of Medical and Life Sciences, Silla University, Busan, 617-736, Republic of Korea

    Han-Seung Lee

  2. Institute of Life Sciences & Resources and Graduate School of Biotechnology, Kyung Hee University, Yongin, 446-701, Republic of Korea

    Hyungjae Lee

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  1. Han-Seung Lee
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  2. Hyungjae Lee
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Correspondence to Hyungjae Lee.

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Lee, HS., Lee, H. Purification and biochemical characterization of bacteriolytic enzyme from Bacillus subtilis YU-1432 active against Porphyromonas gingivalis . J Korean Soc Appl Biol Chem 54, 600–605 (2011). https://doi.org/10.3839/jksabc.2011.090

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  • DOI: https://doi.org/10.3839/jksabc.2011.090

Key words

  • Bacillus subtilis
  • lytic enzyme
  • periodontal disease
  • Porphyromonas gingivalis
  • substrate specificity