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

  • Food Science/Microbiology
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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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References

  • Bollen CM and Quirynen M (1996) Microbiological response to mechanical treatment in combination with adjunctive therapy. A review of the literature. J Periodontol 67, 1143–1158.

    CAS  Google Scholar 

  • Bradford MM (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 72, 248–254.

    CAS  Google Scholar 

  • Choi NS, Chung DM, Han YJ, Kim SH, and Song JJ (2009) Purification and characterization of a subtilisin D5, a fibrinolytic enzyme of Bacillus amyloliquefaciens DJ-5 isolated from doenjang. Food Sci Biotech 18, 500–505.

    CAS  Google Scholar 

  • Dosoretz CG, Chen HC, and Grethlein HE (1990) Effect of environmental conditions on extracellular protease activity in lignolytic cultures of Phanerochaete chrysosporium. Appl Environ Microbiol 56, 395–400.

    CAS  Google Scholar 

  • Fujita M, Nomura K, Hong K, Ito Y, Asada A, and Nishimuro S (1993) Purification and characterization of a strong fibrinolytic enzyme (nattokinase) in the vegetable cheese natto, a popular soybean fermented food in Japan. Biochem Biophys Res Commun 197, 1340–1347.

    CAS  Google Scholar 

  • Hayashi K, Kasumi T, Kubo N, and Tsumura N (1981) Purification and characterization of the lytic enzyme produced by Streptomyces rutgersensis H-46. Agric Biol Chem 45, 2289–2300.

    CAS  Google Scholar 

  • Horz HP and Conrads G (2007) Diagnosis and anti-infective therapy of periodontitis. Expert Rev Anti Infect Ther 5, 703–715.

    CAS  Google Scholar 

  • Jung MH, Kong IS, Bai DH, and Yu JH (1991) Purification and characterization of a bacteriolytic enzyme from alkalophilic Bacillus sp. J Microbiol Biotechnol 1, 102–110.

    CAS  Google Scholar 

  • Jung MH, Ohk SH, Yum DY, Kong IS, Bai DH, and Yu JH (1993) Nucleotide sequence of a bacteriolytic enzyme gene from alkalophilic Bacillus sp. J Microbiol Biotechnol 3, 73–77.

    CAS  Google Scholar 

  • Kim SY, Ohk SH, Bai DH, and Yu JH (1999) Purification and properties of bacteriolytic enzymes from Bacillus licheniformis YS-1005 against Streptococcus mutans. Biosci Biotechnol Biochem 63, 73–77.

    CAS  Google Scholar 

  • Lane DJ (1991) 16S/23S rRNA sequencing. In Nucleic Acid Techniques in Bacterial Systematics, Stackebrandt E and Goodfellow M (eds.), pp.115–175. Wiley & Sons, Chichester, UK.

    Google Scholar 

  • Lee BY, Ohk SH, Kim SY, Bai DH, Yu JH, and Yook SK (1997) Purification of a bacteriolytic enzyme from Bacillus sp. active against Streptococcus mutans. Biotechnol Tech 11, 283–285.

    CAS  Google Scholar 

  • Loesche WJ and Grossman NS (2001) Periodontal disease as a specific, albeit chronic, infection: Diagnosis and treatment. Clin Microbiol Rev 14, 727–752.

    CAS  Google Scholar 

  • Lundblad RL and Noyes CM (1984) In Chemical Reagents for Protein Modification. CRC Press, Boca Raton, FL, USA.

    Google Scholar 

  • Mombelli A and Samaranayake LP (2004) Topical and systemic antibiotics in the management of periodontal diseases. Int Dent J 54, 3–14.

    Google Scholar 

  • Sambrook J and Russell DW (2001) In Molecular Cloning: A Laboratory Manual. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY, USA.

    Google Scholar 

  • Yamazaki K (2007) Relationship between periodontal infections and systemic disease. Clin Microbiol Infect 13 Suppl 4, 3–10.

    Google Scholar 

  • Wang CT, Ji BP, Li B, Nout R, Li PL, Ji H, and Chen LF (2006) Purification and characterization of a fibrinolytic enzyme of Bacillus subtilis DC33, isolated from Chinese traditional Douchi. J Ind Microbiol Biotechnol 33, 750–758.

    CAS  Google Scholar 

  • Wong SL, Price CW, Goldfarb DS, and Doi RH (1984) The subtilisin E gene of Bacillus subtilis is transcribed from a sigma 37 promoter in vivo. Proc Natl Acad Sci USA 81, 1184–1188.

    CAS  Google Scholar 

  • Yu JH (1997) Microbial cell wall lytic enzyme which can be used for industrial and pharmaceutical uses. Foods Biotech 6, 65–66.

    Google Scholar 

  • Yu JH, Jung MH, Park HK, and Bai DH (1992) Cloning and expression in Escherichia coli of a bacteriolytic enzyme gene from alkalophilic Bacillus sp. J Microbiol Biotechnol 2, 161–165.

    CAS  Google Scholar 

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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

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