Mycobiology

The Korean Society of Mycology (한국균학회)
권호 표지
DOI QR코드

DOI QR Code

Immobilization of Keratinase from Aspergillus flavus K-03 for Degradation of Feather Keratin

  • Published : 2005.06.30
Download PDF
⟨ Previous Next ⟩

Abstract

Extracellular keratinase isolated from Aspergillus flavus K-03 was immobilized on calcium alginate. The properties and reaction activities of free and immobilized keratinase with calcium alginate were characterized. The immobilized keratinase showed proteolytic activity against soluble azo-casein and azo-keratin, and insoluble feather keratin. Heat stability and pH tolerance of keratinase were greatly enhanced by immobilization. It also displayed a higher level of heat stability and an increased tolerance toward alkaline pHs compared with free keratinase. During the durability test at $40^{\circ}C$, 48% of the original enzyme activity of the immobilized keratinase was remained after 7 days of incubation. The immobilized keratinase exhibited better stability, thus increasing its potential for use in industrial application.

Keywords

References

  1. Bradford, M. M. 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 https://doi.org/10.1016/0003-2697(76)90527-3
  2. Church, F. C., Swaisgood, H. E. and Catiagnani, G L. 1984. Compositional analysis of proteins following hydrolysis by immobilized proteinases. J. Appl. Biochem. 6: 205-211
  3. Grima, J. P., Chopek, M. K., Titani, K. and Davie, E. W. 1986. Limited proteolysis of human von Willebrand factor by Staphlococcus auren V-8 protease: isolation and partial characterization of a platelet-binding domain. Biochemistry 25: 3156-3163 https://doi.org/10.1021/bi00359a013
  4. Gupta, R., Beg, Q. K. and Lorenz, P. 2002. Bacterial alkaline proteases molecular approaches and industrial applications. Appl. Microbial. Biotechnol. 59: 15-32 https://doi.org/10.1007/s00253-002-0975-y
  5. Janolino, V. G and Swaisgood, H. E. 1982. Analysis and optimization of methods using water-soluble carbodiimide for immobilization of biochemical to porous glass. Biotechnol. Bioeng. 624: 1069-1080
  6. Kim, J.-D. 2003a. Keratinolytic activity of five Aspergillus species isolated from poultry farming soil in Korea. Mycobiology 31: 157-161 https://doi.org/10.4489/MYCO.2003.31.3.157
  7. Kim, J.-D. 2003b. Preliminary characterization of keratinolytic enzyme of Aspergillus flavus K-03 and its potential in biodegradation of keratin wastes. Mycobiology 31: 209-213 https://doi.org/10.4489/MYCO.2003.31.4.209
  8. Lin, X., Shih, J. C. H. and Swaisgood, H. E. 1996. Hydrolysis of feather keratin by immobilized keratinase. Appl. Environ. Microbiol. 62: 4273-4275
  9. Lin, X., Lee, C. G, Casale, E. S. and Shih, J. C. H. 1992. Purification and characterizaton of a keratinase from feather-degrading Bacillus licheniformis strain. Appl. Environ. Microbiol. 58: 3271-3275
  10. Porter, D. H., Swaisgood, H. E. and Gatiagnani, G L. 1984. Characterization of an immobilized digestive enzyme system for determination of protein digestibility. Agric. Food Chem. 32: 334-339 https://doi.org/10.1021/jf00122a038
  11. Rurgess, A. W., Weistein, L. L., Gabel, D. and Scherage, H. A. 1975. Immobilized carboxypeptidase A as a probe for studying the thermally induced unfolding of bovine pancreatic ribonuclease. Biochemistry 14: 197-200 https://doi.org/10.1021/bi00673a001
  12. Swaisgood, H. E. and Catiagnani, G. L. 1987. Use of immobilized proteinases and peptidases to study structural changes in proteins. Methods Enzymol. 135: 596-604 https://doi.org/10.1016/0076-6879(87)35115-8
  13. Swaisgood, H. E. and Catiagnani, G. L. 1991. Protein digestibility. Pp. 309-342. In: Kinsella, J. E. Ed. Advance in food and nutrition research. vol. 35. Elsevir Applied Science Publishers. London
  14. Swaisgood, H. E. and Horton, H. R. 1989. Immobilized enzymes as processing aids or analytical tools. Pp. 242-261. In: Whitaker J. R. and Sonnet, P. E. Eds. ACS Symposium Series 389. American Chemical Society. Washington, DC
  15. Swaisgood, H. E., Chen, S. X. and Catiagnani, G L. 1994. Probing structural changes and preparation of protein domains by proteolysis. Pp. 43-61. In: Yada, R. Y., Jackman, R. L. and Smith, J. L. Eds. Protein structure-function relationship in foods. Blackie Academic and Professional. Glasgow

Cited by

  1. Secretory expression and purification of Bacillus licheniformis keratinase in insect cells vol.12, pp.8, 2017, https://doi.org/10.1371/journal.pone.0183764