Isolation and characterization of a new keratinolytic Bacillus licheniformis strain

  • L. Manczinger
  • M. Rozs
  • Cs. Vágvölgyi
  • F. Kevei


A keratin-degrading bacterium strain (K-508) was isolated from partially degraded feathers and characterized. This isolate exhibited a high chicken feather-degrading activity when cultured in feather-containing broth with a growth optimum of pH 7.0 and 47 °C. On the basis of its phenotypic characteristics (quickly moving, Gram-positive rods), the results of metabolic tests and rDNA sequence analysis, it was identified as Bacillus licheniformis. Its fermentation broth showed activity on N-Bz-l-Phe-l-Val-l-Arg-p-nitroanilide, N-Suc-l-Ala-l-Ala-l-Pro-l-Phe-p-nitroanilide, N-CBZ-Gly-Gly-l-Leu-p-nitroanilide and N-CBZ-l-Ala-l-Ala-l-Leu-p-nitroanilide as chromogenic protease substrates at near neutral pH. Both trypsin-like and chymotrypsin-like proteases were constitutively secreted by this strain.

Bacillus licheniformis chymotrypsin-like protease trypsin-like protease 


Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.


  1. Bierbaum, G., Karutz, M., Weuster-Botz, D. & Wandrey, C. 1994 Production of protease with Bacillus licheniformis mutants insensitive to repression of exoenzyme biosynthesis. Applied Microbiology and Biotechnology 40, 611-617.Google Scholar
  2. Cheng, S.-W., Hu, H.-M., Shen, S.-W., Takagi, H., Asano, M. & Tsai Y.-C. 1995 Production and characterization of keratinase of a feather-degrading Bacillus licheniformis PWD-1. Bioscience Biotechnology and Biochemistry 59, 2239-2243.Google Scholar
  3. Claus, D. & Berkeley, R.C.W. 1986 Genus Bacillus. In Bergey's Manual of Systematic Bacteriology Volume 2. eds. Sneath, P.H.A., Mair, N.S. & Sharpe, M.E. pp. 1105-1139. Baltimore: Williams & Wilkins. ISBN 0-683-07893-3.Google Scholar
  4. El-Naghy, M.A., El-Ktatny, M.S., Fadl-Allah, E.M. & Nazeer, W.W. 1998 Degradation of chicken feathers by Chrysosporium georgiae. Mycopathologia 143, 77-84.Google Scholar
  5. Evans, K.L., Crowder, J. & Miller, E.S. 2000 Subtilisins of Bacillus spp. hydrolyze keratin and allow growth on feathers. Canadian Journal of Microbiology 6, 1004-1011.Google Scholar
  6. Ferrero, M.A., Castro, G.R., Abate, C.M., Baigori, M.D. & Sineriz, F. 1996 Thermostable alkaline proteases of Bacillus licheniformis MIR 29: isolation, production and characterization. Applied Microbiology and Biotechnology 45, 327-332.Google Scholar
  7. Garcia-Kirchner, O., Bautista-Ramirez, M.E. & Segura-Granados, M. 1998 Submerged culture screening of two strains of Streptomyces sp. with high keratinolytic activity. Applied Biochemistry and Biotechnology 70, 277-284.Google Scholar
  8. Gradisar, H., Kern. S. & Friedrich, J. 2000 Keratinase of Doratomyces microsporus. Applied Microbiology and Biotechnology 53, 196-200.Google Scholar
  9. Ignatova, Z., Gousterova, A., Spassov, G. & Nedkov, P. 1999 Isolation and partial characterisation of extracellular keratinase from a wool degrading thermophilic actinomycete strain Thermoactinomyces candidus. Canadian Journal of Microbiology 45, 217-222.Google Scholar
  10. Kaul, S. & Sumbali, G. 1999 Production of extracellular keratinases by keratinophilic fungal species inhabiting feathers of living poultry birds (Gallus domesticus): a comparison. Mycopathologia 146, 19-24.Google Scholar
  11. Lin, X., Inglis, G.D., Yanke, L.J. & Cheng, K.J. 1999 Selection and characterization of feather-degrading bacteria from canola meal compost. Journal of Industrial Microbiology and Biotechnology 23, 149-153.Google Scholar
  12. Lin, X., Kelemen, D.W., Miller, E.S. & Shih, J.C.H. 1995 Nucleotide sequence and expression of kerA, the gene encoding a keratinolytic protease of Bacillus licheniformis PWD-1. Applied and Environmental Microbiology 61, 1469-1474.Google Scholar
  13. Lin, X., Lee, C.-G., Casale, E.S. & Shih, J.C.H. 1992 Purification and characterization of a keratinase from a feather-degrading Bacillus licheniformis strain. Applied and Environmental Microbiology 58, 3271-3275.Google Scholar
  14. Lin, X., Wong, S.-L., Miller, E.S. & Shih, J.C.H. 1997 Expression of the Bacillus licheniformis PWD-1 keratinase gene in B.subtilis. Journal of Industrial Microbiology and Biotechnology 19, 134-138.Google Scholar
  15. Noval, J. & Nickerson, W.J. 1959 Decomposition of native keratin by Streptomyces fradiae. Journal of Bacteriology 77, 73-86.Google Scholar
  16. Onifade, A.A., Al-Sane, N.A., Al-Mussallam, A.A. & Al-Zarbam, S. 1998 Potentials for biotechnological applications of keratin-degrading microorganisms and their enzymes for nutritional improvement of feathers and other keratins as livestock feed resources. Bioresource Technology 66, 1-11.Google Scholar
  17. Papadopoulos, M.C. 1985 Processed chicken feathers as feedstuff for poultry and swine. Agricultural Wastes 14, 275-290.Google Scholar
  18. Papadopoulos, M.C., El Boushy, A.R. & Roodbeen, A.E. 1985 The effect of varying autoclaving conditions and added sodium hydroxide on amino acid content and nitrogen characteristics of feather meal. Journal of Science of Food and Agriculture 36, 1219-1226.Google Scholar
  19. Papadopoulos, M.C., El Boushy, A.R., Roodbeen, A.E. & Ketelaars, E.H. 1986 Effects of processing time and moisture content on amino acid composition and nitrogen characteristics of feather meat. Animal Feed Science Technology 14, 279-290.Google Scholar
  20. Sambrook, J., Fritsch, E.F. & Maniatis, T. 1989 Molecular Cloning: A Laboratory Manual, 2nd ed. Cold Spring Harbor, NY: Cold Spring Harbor Laboratory. ISBN 0-87969-309-6.Google Scholar
  21. Sangali, S. & Brandelli, A. 2000 Feather keratin hydrolysis by a Vibrio sp. strain kr2. Journal of Applied Microbiology 89, 735-743.Google Scholar
  22. Steiner, R.J., Kellems, R.O. & Church, D.C. 1983 Feather and hair meals for ruminants. IV. Effects of chemical treatments of feather and processing time on digestibility. Journal of Animal Science 57, 495-502.Google Scholar
  23. Suzuki, M.T. & Giovannoni, S.J. 1996 Bias caused by template annealing in the amplification of mixtures of 16S rRNA genes by PCR. Applied and Environmental Microbiology 62, 625-630.Google Scholar
  24. Weisburg, W.G., Barns, M., Pelletier, D.A. & Lane, D.J. 1991 Ribosomal DNA amplification for phylogenetic study. Journal of Bacteriology 173, 697-984.Google Scholar
  25. Williams, C.M., Richester, C.S., Mackenzi, J.M. & Shih, J.C.H. 1990 Isolation, identification and characterization of a feather-degrading bacterium. Applied and Environmental Microbiology 56, 1509-1515.Google Scholar

Copyright information

© Kluwer Academic Publishers 2003

Authors and Affiliations

  • L. Manczinger
    • 1
  • M. Rozs
    • 1
  • Cs. Vágvölgyi
    • 1
  • F. Kevei
    • 1
  1. 1.Faculty of Sciences, Department of MicrobiologyUniversity of Szeged

Personalised recommendations