A Thermostable Protease Produced from Bacillus sp. DF 218

Bacillus sp. DF218이 생산하는 내열성 단백질 분해효소

  • Published : 2004.02.28

Abstract

Microorganism (strain DF 218) producing thermostable pretense was isolated from Korean soil and compost. It was Gram-positive, rod-shaped, aerobic, and spore-forming with yellowish white colony color, Temperature range for growth at pH 6.5 was $30-65^{\circ}C$, with optimum growth at $60^{\circ}C$. pH range for growth at $60^{\circ}C$ was 5-7 with optimum of 6.5, which indicates strain DF 218 to be thermophilic. The 16S rDNA sequence of strain DF 218 had 95% sequence similarity with that of Bacillus flexus. Based on physiological properties and phylogenetic analysis, we proposed the isolated strain as Bacillus sp. DF 218. Pretense was produced aerobically at $60^{\circ}C$ for 32 hr in a medium (pH 6.5) containing 1% each trypton, glucose, and NaCl. Its molecular weight was estimated as 61 kDa, with optimum temperature and pH of $60^{\circ}C$ and 7.5, respectively.

Keywords

thermostable microorganism;protease

References

  1. Kageyama K. Studies on Aspergillus oryzae strain for sake brewing. J. Ferment. Technol. 33: 53-57 (1955)
  2. Nunokawa Y, Namba Y, Watanabe S. A study of the rice koji protease. J. Soc. Brew. 53: 930-933 (1961)
  3. Horikoshi K. Production of alkaline enzymes by alkalophilic microorganisims. Part I. Alkaline protease produced by Bacillus sp. No. 221. Agric. Biol. Chem. 35: 1407-1414 (1971) https://doi.org/10.1271/bbb1961.35.1407
  4. Durham DR, Stewart DB, Stellwag EJ. Novel alkaline and heat stable serine protease from alkalophilic Bacillus sp. strain GX6638. J. Bacteriol. 169: 2762-2768 (1987) https://doi.org/10.1128/jb.169.6.2762-2768.1987
  5. Michels PC, Douglass C. Pressure-enhanced activity and stability of a hyperthermophilic protease from a deep-sea methanogen. Appl. Environ. Microbiol. 63: 3985-3991 (1997)
  6. Vazquez D, Cantera MB. A simple and rapid technique for postelectrophoretic detection of protease using azocasein. Electrophoresis 16: 1894-1897 (1995) https://doi.org/10.1002/elps.11501601311
  7. Tasi Y, Juang R, Lin S, Chen S, Yamasaki M, Tamura G. Production and further characterization of an alkalophilic elastase produced by alkalophilic Bacillus strain Ya-B. Appl. Environ. Microbiol. 54: 3156-3161 (1998)
  8. Massaki Y, Kazuo S, Mitsuo M. Purification and properties of acid protease from Monascus sp. No. 3405. Agric. Biol. Chem. 48: 1637-1645 (1984) https://doi.org/10.1271/bbb1961.48.1637
  9. Ward OP. Proteolytic enzymes, Vol. III, pp. 789-818. In: Comprehensive Biotechnology. Elsevier, New York, NY, USA (1986)
  10. Davis LG, Dibner MD, Battey JF. Basic Method in Molecular Biology. Elsevier, New York, NY, USA. p. 42 (1986)
  11. Kim HK, Kim KH, Lee JK, Kim YO, Nam HS, Oh TK. Characterization of a thermostable protease from thermophilic Bacillus amyloliquefaciens NS 15-4. Korean J. Appl. Microbiol. Biotechnol. 23: 322-328 (1995)
  12. MacFaddin JF. Biochemical Tests for Identification of Medical Bacteria. 2nd ed. Williams & Wilkins, Baltimore, MD, USA (1990)
  13. Priest FG, Goodfellow M, Todd C. A numerical classification of the genus Bacillus. J. Gen. Microbiol. 134: 1847-1882 (1988)
  14. Kobayashi T, Ogasawara A, Ito S, Saitoh M. Purification and some properties of alkaline proteinase produced by Pseudomonas maltophilia. Agric. Biol. Chem. 49: 693-698 (1985) https://doi.org/10.1271/bbb1961.49.693
  15. Banerjee UC, Sani RK, Azimi W, Soni R. Thermostable alkaline protease from Bacillus brevis and its characterization as a laundry detergent additive. Process Biochem. 30: 213-219 (1999)
  16. Peckman EV. Aspergillus proteinase. Biochemistry 5: 321-325 (1951)
  17. Beynon RJ, Bond JS. Proteolytic Enzymes-a Practical Approach, IRL press, Oxford, UK (1989)
  18. Crewth WC. The effect of pH and cations on the thermal denaturation of trypsin. J. Biol. Chem. 6: 597-601 (1963)
  19. Nakadai T, Nasuno S, Iguchi N. Purification and some properties of alkaline proteinase from Aspergillus oryzae. Agric. Biol. Chem. 37: 2685-2694 (1973) https://doi.org/10.1271/bbb1961.37.2685
  20. Kreig NR, Halt JG. Bergey's Manual of Systematic Bacteriology. Williams and Wilkins Co., Baltimore, MD, USA (1984)