Abstract
A keratinolytic enzyme (KerA1) secreted by a newly isolated Bacillus pumilus strain A1 cultivated in medium containing chicken feather meal was purified and characterized, and the gene was isolated and sequenced. The molecular mass of the purified enzyme was estimated to be 34,000 Da by sodium dodecyl sulfate (SDS) polyacrylamide gel electrophoresis and gel filtration. The optimum pH and temperature for the purified keratinase were 9.0 and 60 °C, respectively, using keratin as a substrate. KerA1 showed a high stability towards nonionic surfactants. It was found to be relatively stable toward the strong anionic surfactant (SDS). The deduced amino acid sequence of the keratinase KerA1 differs from both the organic solvent tolerant protease of B. pumilus 115b and the dehairing protease of B. pumilus UN-31-C-42 by one and nine amino acids, respectively. These results suggest that this keratinase may be a useful alternative and ecofriendly route for handling the abundant amount of waste feathers and for applications in detergent formulations.
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Onifade, A., Al-Sane, N. A., Al-Musalam, A. A., & Al-Zarban, S. (1998). Bioresource Technology, 66, 1–11.
Gupta, R., & Ramnani, P. (2006). Applied Microbiology and Biotechnology, 70, 21–33.
Fakhfakh, N., Kanoun, S., Manni, L., & Nasri, M. (2009). Canadian Journal of Microbiology, 55, 427–436.
Kumar, C. G. (2002). Letters in Applied Microbiology, 34, 13–17.
Williams, C. M., Lee, C. G., Garlich, J. D., & Shih, J. C. H. (1991). Poultry Science, 70, 85–94.
Odetallah, N. H., Wang, J. J., Garlich, J. D., & Shih, J. C. (2003). Poultry Science, 82, 664–670.
Langeveld, J. P., Wang, J. J., Van de Wiel, D. F., Shih, G. C., Garssen, G. J., Bossers, A., et al. (2003). Journal of Infectious Diseases, 188, 1782–1789.
Huang, Q., Peng, Y., Li, X., Wang, H., & Zhang, Y. (2003). Current Microbiology, 46, 169–173.
Anbu, P., Gopinath, S. C. B., Hilda, A., Lakshmi-Priya, T., & Annadurai, G. (2005). Enzyme and Microbial Technology, 36, 639–647.
Suh, H. J., & Lee, H. K. (2001). Journal of Protein Chemistry, 20, 165–169.
Cai, Cg, Chen, Js, Qi, Jj, Yin, Y., & Zheng, Xd. (2008). Journal of Zhejiang University. Science B, 9, 713–720.
Bressollier, P., Letourneau, F., Urdaci, M., & Verneuil, B. (1999). Applied and Environmental Microbiology, 65, 2570–2576.
Tatineni, R., Doddapaneni, K. K., Potumarthi, R. C., Vellanki, R. N., Kandathil, M. T., Kolli, N., et al. (2008). Bioresource Technology, 99, 1596–1602.
Syed, D. G., Lee, J. C., Li, W. J., Kim, C. J., & Agasar, D. (2009). Bioresource Technology, 100, 1868–1871.
Takiuchi, I., Higuchi, D., Sei, Y., & Koga, M. (1982). Sabouraudia, 20, 281–288.
Kembhavi, A. A., Kulkarni, A., & Pant, A. (1993). Applied Biochemistry and Biotechnology, 38, 83–92.
Bradford, M. (1976). Analytical Biochemistry, 72, 248–254.
Laemmli, U. K. (1970). Nature, 227, 680–685.
Garcia-Carreno, F. L., Dimes, L. E., & Haard, N. F. (1993). Analytical Biochemistry, 214, 65–69.
Tsai, I. H., Chuang, K. L., & Chuang, J. L. (1986). Biochemistry and Molecular Biology, 85, 235–239.
Pan, J., Huang, Q., & Zhang, Y. (2004). Current Microbiology, 49, 165–169.
Haddar, A., Bougatef, A., Agrebi, R., Sellami-Kamoun, A., & Nasri, M. (2009). Process Biochemistry, 44, 29–35.
Pillai, P., & Archana, G. (2008). Applied Microbiology and Biotechnology, 78, 643–650.
Han, X. Q., & Damodaran, S. (1998). Journal of Agricultural and Food Chemistry, 46, 3596–3603.
Miyaji, T., Otta, Y., Nakagawa, T., Watanabe, T., Niimura, Y., & Tomizuka, N. (2006). Letters in Applied Microbiology, 42, 242–247.
Lin, X., Shih, J. C. H., & Swaisgood, H. E. (1996). Applied and Environmental Microbiology, 62, 4273–4275.
Radha, S., & Gunasekaran, P. (2008). Bioresource Technology, 99, 5528–5537.
Manni, L., Jellouli, K., Agrebi, R., Bayoudh, A., & Nasri, M. (2008). Process Biochemistry, 43, 522–530.
Kidd, R. D., Yennawar, H. P., Sears, P., Wong, C. H., & Farber, G. K. (1996). Journal of the American Chemical Society, 118, 1645–1650.
Smith, C. A., Toogood, H. S., Baker, H. M., Daniel, R. M., & Baker, E. N. (1999). Journal of Molecular Biology, 294, 1027–1040.
Lin, X., Lee, C. G., Casale, E. S., & Shih, J. C. H. (1992). Applied and Environmental Microbiology, 58, 3271–3275.
Haddar, A., Agrebi, R., Bougatef, A., Hmidet, N., Sellami-Kamoun, A., & Nasri, M. (2009). Bioresource Technology, 100, 3366–3373.
Evans, K. L., Crowder, J. S., & Miller, E. S. (2000). Canadian Journal of Microbiology, 46, 1004–1011.
Mignon, B., Swinnen, M., Bouchara, J. P., Hofinger, M., Nikkels, A., Pierard, G., et al. (1998). Medical Mycology, 36, 395–404.
Gioia, J., Yerrapragada, S., Qin, X., Jiang, H., Igboeli, O. C., Muzny, D., et al. (2007). SAFR-032 PLoS ONE, 2, e928.
Abd Rahman, R. N. Z. R., Mahamad, S., Salleh, A. B., & Basri, M. (2007). Journal of Industrial Microbiology & Biotechnology, 34, 509–517.
Acknowledgment
This work was funded by “Ministry of Higher Education, Scientific Research and Technology-Tunisia”. The authors would like to thank Mr. A. Hajji from the Faculty Letters and Human Sciences of Kairouan-Tunisia for his help with the English language.
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Fakhfakh-Zouari, N., Hmidet, N., Haddar, A. et al. A Novel Serine Metallokeratinase from a Newly Isolated Bacillus pumilus A1 Grown on Chicken Feather Meal: Biochemical and Molecular Characterization. Appl Biochem Biotechnol 162, 329–344 (2010). https://doi.org/10.1007/s12010-009-8774-x
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DOI: https://doi.org/10.1007/s12010-009-8774-x