Abstract
An α-keratinase producing strain was isolated with wool as the sole carbon and nitrogen source and identified as Bacillus pumilus K9. The major amino acids liberated from the keratin degradation of wool by B. pumilus K9 were glutamic acid and leucine. The α-keratinase was purified to electrophoretic homogeneity with a molecular weight of 32000. The purified enzyme exhibits an optimum activity at 60 °C and pH=9.0. It was stable at pH values between 8 and 11. Bacillas pumilus keratinase displays a high activity towards casein, keratin, wool and feather, which indicates its wide application range. The keratinase was completely inhibited by phenylmethyl-sulfonyl fluoride(PMSF) and β-mercaptoethanol, and moderately inhibited by ethylemediamine-tetraacetic acid(EDTA), suggesting it is a metallo-cysteine keratinase. This enzyme could remain stable that could even be promoted in the presence of surfactants, including sodium dodecyl sulfate(SDS), Tween and Triton. And Tween 40 and Triton X-100 could substantially enhance the activity of the enzyme by 54% and 35%, respectively. It may indicate the prominent feature of the keratinase to tolerate surfactants. The enzymatic properties distinguish this keratinase from others in the literature. Furthermore, this enzyme is extremely stable in the presence of a commercially available detergent with 1% concentration. Detergents ARIEL, Bluemoon and WhiteCat can enhance the activity of the keratinase by 43.56%, 15.22%, and 22.48%, respectively.
Similar content being viewed by others
References
Riffel A., Brandelli A., Bellato C. M., Souza G. H., Eberlin M. N., Tavares F. C., J. Biotechnol., 2007, 128, 693
Gupta R., Sharma R., Beg Q. K., Crit. Rev. Biotechnol., 2013, 33, 216
Brandelli A., Daroit D. J., Riffel A., Appl. Microbiol. Biotechnol., 2010, 85, 1735
Anbu P., Gopinatha S. C. B., Hildaa A., Lakshmi P. T., Annaduraic G., Enzyme Microb. Technol., 2005, 36, 639
Syed D. G., Lee J. C., Li W. J., Kim C. J., Agasar D., Bioresour. Technol., 2009, 100, 1868
Farag A. M., Hassan M. A. Enzyme Microb. Technol., 2004, 34, 85
Lin X., Lee C. G., Casale E. S., Shih J. C., Appl. Environ. Microbiol., 1992, 58, 3271
Bressollier P., Letourneau F., Urdaci M., Verneuil B., Appl. Environ. Microbiol., 1999, 65, 2570
Cassano A., Drioli E., Molinari R., Grimaldi D., La C. F., Rossi M., J. Soc. Leather Technol. Chem., 2000, 84, 205
Abdel S., Manal M., Heba I., Nabil H., Mohamed H., Int. Biodeter. Biodegr., 2012, 70, 14
Kumar S., Tamura K., Nei M., Brief. Bioinform., 2004, 5, 150
Bradford M. M., Anal. Biochem., 1976, 72, 248
Yamamura S., Morita Y., Hasan Q., Rao S. R., Murakami Y., Yokoyama K., Tamiya E., J. Biosci. Bioeng., 2002, 93, 595
Laemmli U. K. Nature, 1970, 227, 680
Fakhfakh-Zouari N., Hmidet N., Haddar A., Kanoun S., Nasri M., Appl. Biochem. Biotechnol., 2010, 162, 329
Suntornsuk W., Tongjun J., Onnim P., Oyama H., Ratanakanokchai K., Kusamran T., Oda K., World J. Microbiol. Biotechnol., 2005, 21, 1111
Mitsuiki S., Ichikawa M., Oka T., Sakai M., Moriyama Y., Sameshma Y., Goto M., Furukawa K., Enzyme Microb. Technol., 2004, 34, 482
Bernal C., Cairo J., Coello N., Enzyme Microb. Technol., 2006, 38, 49
Tatineni R., Doddapaneni K. K., Potumarthi R. C., Vellanki R. N., Kandathil M. T., Kolli N., Mangamoori L. N., Bioresource Technol., 2008, 99, 1596
Cavello I. A., Hours R. A., Rojas N. L., Cavalitto S. F., Process Biochem., 2013, 48, 972
Lv L. X., Sim M. H., Li Y. D., Min J., Feng W. H., Guan W. J., Li Y. Q., Process Biochem., 2010, 45, 1236
Correa A. P., Daroit D. J., Brandelli A., Int. Biodeter. Biodegr., 2010, 64, 1
Bernal C., Vidal L., Valdivieso E., Coello N., World J. Microbiol. Biotechnol., 2003, 19, 255
Li X., Dong H., Zhang H., Chem. J. Chinese Universities, 2013, 34(12), 2861
Cao L., Tan H., Liu Y., Xue X., Zhou S., Lett. Appl. Microbiol., 2008, 46, 389
Chen K. N., Huang J. C., Chung C. I., Kuo W. Y., Chen M. J., World J. Microbiol. Biotechnol., 2011, 27, 349
Santos R. M. D. B., Firmino A. A., de Sá C. M., Felix C. R., Curr. Microbiol., 1996, 33, 364
Gradisar H., Friedrich J., Krizaj I., Jerala R., Appl. Environ. Microbiol., 2005, 71, 3420
Paul T., Das A., Mandal A., J. Clean. Prod., 2014, 66, 672
Rajput R., Sharma R., Gupta R., Enzyme Res., 2010, 2010, 132148
Aravindhan R., Saravanabhavan S., Rao J. R., Nair B. U., J. Am. Leather Chem. As., 2004, 992, 53
Author information
Authors and Affiliations
Corresponding author
Additional information
Supported by the National High Technology Research and Development Program of China(No.2012AA022204C) and the National Natural Science Foundation of China(No.21406088).
Rights and permissions
About this article
Cite this article
Gong, J., Wang, Y., Zhang, D. et al. A surfactant-stable Bacillus pumilus K9 α-keratinase and its potential application in detergent industry. Chem. Res. Chin. Univ. 31, 91–97 (2015). https://doi.org/10.1007/s40242-015-4351-8
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s40242-015-4351-8