Skip to main content
SpringerLink
Log in

Characterization of a novel Stenotrophomonas isolate with high keratinase activity and purification of the enzyme

  • Original Paper
  • Published:
Journal of Industrial Microbiology & Biotechnology

Abstract

A feather-degrading bacterium was isolated from poultry decomposition feathers in China. The strain, named L1, showed significant feather-degrading activity because it grew and reproduced quickly on basal medium containing 10 g/L of native feather as the source of energy, carbon, and nitrogen. According to the phenotypic characteristics and 16S rRNA profile, the isolate belongs to Stenotrophomonas maltophilia. Keratinase activity of the isolate was determined during cultivation on raw feathers at different temperatures and initial pH. Maximum growth and feather-degrading activity of the bacterium were observed at 40°C and initial pH ranging from 7.5 to 8.0. The crude enzyme was purified by ammonium sulphate precipitation, Sephadex G-100 chromatographic and ceramic hydroxyapatite (CHT) chromatographic. Its molecular mass estimated as 35.2 kDa in SDS-PAGE. The enzyme had an optimum activity at the pH was 7.8 and the temperature was 40°C. The keratinase was wholly inhibited by a serine protease inhibitor, PMSF. Its activity was activated or inhibited by different metal ions. The keratinase activity of enzyme from strain L1 functioned on different keratins, such as feather, hair, wool, horn, and so on.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6
Fig. 7
Fig. 8

Similar content being viewed by others

References

  1. Williams CM, Lee CG, Garlich JD, Shih JCH (1991) Evaluation of a bacterial feather fermentation product, feather-lysate, as a feed protein. Poultry Sci 70:85–94

    CAS  Google Scholar 

  2. Gupta R, Ramnani P (2006) Microbial keratinases and their prospective applications: an overview. Appl Microbiol Biotechnol 70:21–33

    Article  PubMed  CAS  Google Scholar 

  3. Onifade AA, Al-Sane NA, Al-Musallam AA, Al-Zarban S (1998) A review: potentials for biotechnological applications of keratin-degrading microorganisms and their enzymes for nutritional improvement of feathers and other keratins as livestock feed resources. Bioresour Technol 66:1–11

    Article  CAS  Google Scholar 

  4. Zaghloul TI, Haroun MA, El-Gayar K, Abdelal A (2004) Recycling of keratin-containing materials (chicken feather) through genetically engineered bacteria. Polym Plast Technol Eng 43:1589–1599

    Article  CAS  Google Scholar 

  5. Lee CG, Ferket PR, Shih JCH (1991) Improvement of feather digestibility by bacterial keratinase as a feed additive. FASEB J 5:596–596

    Google Scholar 

  6. Langeveld JPM, Wang JJ, Van de Wiel DFM, Shih GC, Garssen GJ, Bossers A, Shih JCH (2003) Enzymatic degradation of prion protein in brain stem from infected cattle and sheep. J Infect Dis 188:1782–1789

    Article  PubMed  CAS  Google Scholar 

  7. Mitsuiki S, Hui Z, Matsumoto D, Sakai M, Moriyama Y, Furukawa K, Kanouchi H, Oka T (2006) Degradation of PrPSc by keratinolytic protease from Nocardiopsis sp TOA-1. Biosci Biotechnol Biochem 70:1246–1248

    Article  PubMed  CAS  Google Scholar 

  8. Wang JH, Borwornpinyo R, Odetallah N, Shih JCH (2005) Enzymatic degradation of a prion-like protein, Sup35NM-His6. Enzyme Microb Technol 36:758–765

    Article  CAS  Google Scholar 

  9. Yoshioka M, Miwa T, Horii H, Takata M, Yokoyama T, Nishizawa K, Watanabe M, Shinagawa M, Murayama Y (2007) Characterization of a proteolytic enzyme derived from a Bacillus strain that effectively degrades prion protein. J Appl Microbiol 102:509–515

    Article  PubMed  CAS  Google Scholar 

  10. Gradisar H, Kern S, Friedrich J (2000) Keratinase of Doratomyces microsporus. Appl Microbiol Biotechnol 53:196–200

    Article  PubMed  CAS  Google Scholar 

  11. Kumar S, Tamura K, Nei M (2004) MEGA3: Integrated software for Molecular Evolutionary Genetics Analysis and sequence alignment. Brief Bioinform 5:150–163

    Article  PubMed  CAS  Google Scholar 

  12. 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

    Article  PubMed  CAS  Google Scholar 

  13. Laemmli UK (1970) Cleavage of Structural Proteins during the Assembly of the Head of Bacteriophage T4. Nature 227:680–685

    Article  PubMed  CAS  Google Scholar 

  14. Bockle B, Galunsky B, Muller R (1995) Characterization of a keratinolytic serine proteinase from Streptomyces pactum DSM 40530. Appl Environ Microbiol 61:3705–3710

    PubMed  CAS  Google Scholar 

  15. Kim JM, Lim WJ, Suh HJ (2001) Feather-degrading Bacillus species from poultry waste. Process Biochem 37:287–291

    Article  CAS  Google Scholar 

  16. Zaghloul Ti (1998) Cloned Bacillus subtilis alkaline protease (aprA) gene showing high level of keratinolytic activity. Appl Biochem Biotechnol 70–2:199–205

    Article  Google Scholar 

  17. Riffel A, Brandelli A (2002) Isolation and characterization of a feather-degrading bacterium from the poultry processing industry. J Ind Microbiol Biotechnol 29:255–258

    Article  PubMed  CAS  Google Scholar 

  18. Lucas FS, Broennimann O, Febbraro I, Heeb P (2003) High diversity among feather-degrading bacteria from a dry meadow soil. Microb Ecol 45:282–290

    Article  PubMed  CAS  Google Scholar 

  19. Sangali S, Brandelli A (2000) Isolation and characterization of a novel feather-degrading bacterial strain. Appl Biochem Biotechnol 87:17–24

    Article  PubMed  CAS  Google Scholar 

  20. Yamamura S, Morita Y, Hasan Q, Rao SR, Murakami Y, Yokoyama K, Tamiya E (2002) Characterization of a new keratin-degrading bacterium isolated from deer fur. J Biosci Bioeng 93:595–600

    PubMed  CAS  Google Scholar 

  21. Yamamura S, Morita Y, Hasan Q, Yokoyama K, Tamiya E (2002) Keratin degradation: a cooperative action of two enzymes from Stenotrophomonas sp. Biochem Biophys Res Commun 294:1138–1143

    Article  PubMed  CAS  Google Scholar 

  22. Suntornsuk W, Tongjun J, Onnim P, Oyama H, Ratanakanokchai K, Kusamran T, Oda K (2005) Purification and characterisation of keratinase from a thermotolerant feather-degrading bacterium. World J Microbiol Biotechnol 21:1111–1117

    Article  CAS  Google Scholar 

  23. Bressollier P, Letourneau F, Urdaci M, Verneuil B (1999) Purification and characterization of a keratinolytic serine proteinase from Streptomyces albidoflavus. Appl Environ Microbiol 65:2570–2576

    PubMed  CAS  Google Scholar 

  24. Friedrich AB, Antranikian G (1996) Keratin Degradation by Fervidobacterium pennavorans, a novel thermophilic anaerobic species of the order thermotogales. Appl Environ Microbiol 62:2875–2882

    PubMed  CAS  Google Scholar 

  25. Gradisar H, Friedrich J, Krizaj I, Jerala R (2005) Similarities and specificities of fungal keratinolytic proteases: comparison of keratinases of Paecilomyces marquandii and Doratomyces microsporus to some known proteases. Appl Environ Microbiol 71:3420–3426

    Article  PubMed  CAS  Google Scholar 

Download references

Acknowledgments

This work was partially supported by Program for Changjiang Scholars and Innovative Research Team in University (No. IRT0526) and partially financed by Shanghai Leading Academic Discipline Project (No. B604).

Author information

Authors and Affiliations

  1. Key Laboratory of Science and Technology of Eco-Textile, Donghua University, Ministry of Education, 201620, Shanghai, China

    Zhang-Jun Cao, Dong-Kai Wei, Xing-Qun Zhang & Mei-Hua Zhou

  2. College of Chemistry, Chemical Engineering and Biotechnology, Donghua University, 201620, Shanghai, China

    Zhang-Jun Cao, Dong-Kai Wei & Xing-Qun Zhang

  3. College of Environmental Science and Engineering, Donghua University, 201620, Shanghai, China

    Qi Zhang, Li Chen, Jing Wang & Mei-Hua Zhou

Authors
  1. Zhang-Jun Cao
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Qi Zhang
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Dong-Kai Wei
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Li Chen
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Jing Wang
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Xing-Qun Zhang
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Mei-Hua Zhou
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Mei-Hua Zhou.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Cao, ZJ., Zhang, Q., Wei, DK. et al. Characterization of a novel Stenotrophomonas isolate with high keratinase activity and purification of the enzyme. J Ind Microbiol Biotechnol 36, 181–188 (2009). https://doi.org/10.1007/s10295-008-0469-8

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s10295-008-0469-8

Keywords

Search

Navigation