Hide depilation and feather disintegration studies with keratinolytic serine protease from a novel Bacillus subtilis isolate

Biotechnologically Relevant Enzymes and Proteins


Keratinases play an important role in biotechnological applications such as improvement of feather meal, enzymatic dehairing and production of amino acids or peptides from high molecular weight substrates. Bacillus subtilis P13, isolated from Vajreshwari hot spring (45–50°C) near Mumbai, India, produces a neutral serine protease and has an optimum temperature of 65°C. This enzyme preparation was keratinolytic in nature and could disintegrate whole chicken feathers, except for the remnants of shafts. The enzyme preparation also exhibited depilation of goat hides with the recovery of intact animal hair. The enzyme preparation could release peptides from ground feathers and bring about their weight reduction; however, similar action on hair was relatively weak. A single major PMSF-sensitive protease band could be detected upon zymogram analysis, indicating that a single enzyme may be responsible for feather degradation and hide depilation. The importance of these findings in the biotechnological application for feather and leather industries is discussed.


Keratinase Bacillus subtilis P13 Feather disintegration Hide dehairing 


  1. Böckle B, Müller R (1997) Reduction of disulfide bonds by Streptomyces pactum during growth on chicken feather. Appl Environ Microbiol 63:790–792Google Scholar
  2. Böckle B, Galunsky B, Müller R (1995) Characterization of a keratinolytic serine proteinase from Streptomyces pactum DSM 40530. Appl Environ Microbiol 61:3705–3710Google Scholar
  3. Bressollier P, Letourneau F, Urdaci M, Verneuil B (1999) Purification and characterization of a keratinolytic serine protease from Streptomyces albidoflavus. Appl Environ Microbiol 65:2570–2576Google Scholar
  4. Dozie INS, Okeke CN, Unaeze NC (1994) A thermostable, alkaline-active keratinolytic proteinase from Chrysosporium keratinophilum. World J Microbiol Biotechnol 10:563–567CrossRefGoogle Scholar
  5. Giongo LJ, Lucas SF, Casarin F, Heeb P, Brandelli A (2007) Keratinolytic proteases of Bacillus species isolated from the Amazon basin showing remarkable de-hairing activity. World J Microbiol Biotechnol 23:375–382CrossRefGoogle Scholar
  6. Gousterova A, Brailkova D, Goshev I Christov P, Tishinov K, Tonkova VE, Haertle T, Nedlkov P (2005) Degradation of keratin and collagen containing wastes by newly isolated Thermoactinomycetes for alkaline hydrolysis. Lett Appl Microbiol 40:335–340CrossRefGoogle Scholar
  7. Gradišar H, Friedrich J, Križaj I, Jerala R (2005) Similarities and specificities of fungal keratinolytic proteases: comparison of keratinases of Pacecilomyces marquandii and Doratomyyces microsporus to some known proteases. Appl Environ Microbiol 71:3420–3426CrossRefGoogle Scholar
  8. Gradišar H, Kern S, Friedrich J (2000) Keratinase of Doratomyces microsporus. Appl Microbiol Biotechnol 53:196–200CrossRefGoogle Scholar
  9. Gupta R, Ramnani P (2006) Microbial keratinases and their prospective applications: An overview. Appl Microbiol Biotechnol 70:21–33CrossRefGoogle Scholar
  10. Holt JG, Krieg NR, Sneath PHA, Staley JT (1994) Bergey’s manual of determinative bacteriology, 9th edition. Williams and Wilkins, Baltimore, pp 559–562Google Scholar
  11. Janssen PH, Peek K, Morgan HW (1994) Effect of culture condition on the production of an extracellular proteinase by Thermus Sp Rt 41 A. Appl Microbiol Biotechnol 41:400–406CrossRefGoogle Scholar
  12. Joo HS, Kumar CG, Park GC, Kim KT, Paik SR, Chang CS (2002) Optimization of the production of an extracellular alkaline protease from Bacillus horikoshii. Process Biochem 38:155–159CrossRefGoogle Scholar
  13. Kato T, Yamagata Y, Aria T, Ichishima E (1992) Purification of a new extracellular 90-kDa serine proteinase with isoelectric point 3.9 from Bacillus subtilis nattoo and elucidation of its distinct mode of action. Biosci Biotechnol Biochem 56:1166–1168CrossRefGoogle Scholar
  14. Lin X, Lee CG, Casale ES, Shih JCH (1992) Purification and characterization of a keratinase from a feather—degrading Bacillus licheniformis strain. Appl Environ Microbiol 58:3271–3275Google Scholar
  15. Macedo AJ, da Silva WOBD, Gava R, Driemeier D, Henriques JAP, Termignoni C (2005) Novel keratinase from Bacillus subtilis S14 exhibiting remarkable dehairing capabilities. Appl Environ Microbiol 71:594–596CrossRefGoogle Scholar
  16. Nam G-W, Lee D-W, Lee H-S, Lee N-J, Kim B-C, Choe E-A, Hwang J-K, Suhartono MT, Pyun Y-R (2002) Native-feather degradation by Fervidobacterium islandicum AW-1, a newly isolated keratinase-producing thermophilic anaerobe. Arch Microbiol 178:538–547CrossRefGoogle Scholar
  17. Onafide AA, Sane NA, Musallam AA, Zarban S (1998) A review: potentials for biotechnological application of keratin-degrading microorganisms and their enzymes for nutritional improvement of feather and other keratins as livestock feed resources. Bioresour Technol 66:1–11CrossRefGoogle Scholar
  18. Papadopoulos MC (1986) The effect of enzymatic treatment on amino acid content and nitrogen characteristics of feather meal. Anim Feed Sci Technol 16:151–156CrossRefGoogle Scholar
  19. Puri S, Beg QK, Gupta R (2002) Optimization of protease production from Bacillus sp. using surface response methodology. Curr Microbiol 44:286–290CrossRefGoogle Scholar
  20. Ramnani P, Singh R, Gupta R (2005) Keratinolytic potential of Bacillus licheniformis RG 1: structural and biochemical mechanism of feather degradation. Can J Microbiol 51:191–196CrossRefGoogle Scholar
  21. Rao MB, Tanksale AM, Ghatge MS Deshpande VV (1998) Molecular and Biotechnological aspects of microbial proteases. Microbiol Mol Biol Rev 62:597–635Google Scholar
  22. Riffel A, Lucas F, Heeb P, Brandelli A (2003) Characterization of a new keratinolytic bacterium that completely degrades native feather keratin. Arch Microbiol 179:258–265Google Scholar
  23. Saravanabhavan S, Aravindhan R, Thanikaivelan P, Rao JR, Nair BU, Ramasami T (2004) A source reduction approach: integrated bio-based tanning methods and role of enzymes in dehairing and fiber opening. Clean Technol Environ Policy 7:3–14CrossRefGoogle Scholar
  24. Sambrook J, Russell DW (2001) Molecular cloning a laboratory manual, 3rd edn. Cold Spring Harbour Laboratory, New YorkGoogle Scholar
  25. Schrooyen PMM, Dijkstra PJ, Oberthur RC, Bantjes A, Feijen J (2001) Partially carboxymethylated feather keratins. 2. Thermal and mechanical properties of films. J Agric Food Chem 49:221–230CrossRefGoogle Scholar
  26. Shivaji S, Bhanu VN, Aggarwal RK (2000) Identification of Yersinia pestis as a causative organism of plague in India as determined by 16S rDNA sequencing and RAPD-based genomic finger printing. FEMS Microbiol Lett 189:247–252CrossRefGoogle Scholar
  27. Suzuki Y, Tsujimoto Y, Matsui H, Watanable K (2006) Decomposition of extremely hard-to-degrade animal proteins by thermophilic bacteria. J Biosci Bioeng 102:73–81CrossRefGoogle Scholar
  28. Thanikavelan P, Rao JR, Nair BU, Ramasami T (2004) Progress and recent trends in biotechnological methods for leather processing. Trends Biotechnol 22:181–186CrossRefGoogle Scholar
  29. Thys RCS, Lucas FS, Riffel A, Heeb P, Brandelli A (2004) Characterization of a protease of a feather—degrading Microbacterium species. Lett Appl Microbiol 39:181–186CrossRefGoogle Scholar
  30. Varela H, Ferrari MD, Belobrajdic L, Vasquez A, Loprena MD (1997) Skin dehairing proteases of Bacillus subtilis: production and partial characterization. Biotechnol Lett 19:755–758CrossRefGoogle Scholar
  31. Voet D, Voet JG (2004) Biochemistry, 3rd edn. Wiley, New YorkGoogle Scholar
  32. Williams CM, Richter CS, Mackenzie JM, Shih JCH (1990) Isolation, identification and characterization of a feather degrading bacterium. Appl Environ Microbiol 56:1509–1515Google Scholar
  33. Yamagata Y, Abe R, Fugita Y, Ichishima E (1995) Molecular cloning and nucleotide sequence of the 90k serine protease gene hspK from Bacillus subtilis (natto) No. 16. Curr Microbiol 31:340–344CrossRefGoogle Scholar
  34. Yamamura S, Morita Y, Hasan Q, Yokoyama K, Tamiya E (2002) Keratin degradation: a co-operative action of two enzymes from Stenotrophomonas sp. Biochem Biophys Res Commun 294:1138–1143CrossRefGoogle Scholar

Copyright information

© Springer-Verlag 2008

Authors and Affiliations

  1. 1.Department of Microbiology and Biotechnology Centre, Faculty of ScienceThe Maharaja Sayajirao University of BarodaVadodaraIndia

Personalised recommendations