Isolation and Characterization of Proteases Enzyme from Locally Isolated Bacillus sp.
American Journal of Life Sciences
Volume 2, Issue 6, December 2014, Pages: 338-344
Received: Nov. 7, 2014; Accepted: Nov. 15, 2014; Published: Nov. 20, 2014
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Authors
Md. Ekhlas Uddin, Dept. Of Biotechnology & Genetic Engineering, Islamic University, Kushtia, Bangladesh
Pulak Maitra, Dept. Of Biotechnology & Genetic Engineering, Islamic University, Kushtia, Bangladesh
Hossain Md. Faruquee, Dept. Of Biotechnology & Genetic Engineering, Islamic University, Kushtia, Bangladesh; Vaccine research Laboratory, Gono University, Savar, Dhaka
Md. Firoz Alam, Dept. Of Biotechnology & Genetic Engineering, Islamic University, Kushtia, Bangladesh
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Abstract
A bacterium was isolated from the natural source. Gram staining & spore staining showed that the organism is gram positive and forms spore during adverse condition in the growth medium. After various tests it was suggested and the features agreed with the description of Bacillus sp in Bergey’s Manual of Systematic Bacteriology [24]. It was also identified as Bacillus sp with 99.9% identity by API 50 CHB. In growth curve determination showed that the growth of the organism is increased with the increase of incubation period and the growth reached maximum at around 24 hours of incubation and the protease activity was the maximum of the 26 hours culture. This microbe has grown at high temperature and pressure. Its optimum pH and temperature were 8.5 and 60°C. It secretes an extracellular protease in the growth medium. The enzyme hydrolyses a number of proteins including azocasein which suggests that that it is an extracellular protease. The enzyme seems to be alkaline protease which is capable of De-Hairing from skin and hides. A number of companies such as NOVO chemicals started to produce NOVOzymes for tannery industries. The potential for use of microbial enzymes in leather processing lies mainly in areas in which pollution-causing chemicals are being used.
Keywords
Identification, Characterization of Protease, Bacillus sp., Isolation, Alkaline Protease
To cite this article
Md. Ekhlas Uddin, Pulak Maitra, Hossain Md. Faruquee, Md. Firoz Alam, Isolation and Characterization of Proteases Enzyme from Locally Isolated Bacillus sp., American Journal of Life Sciences. Vol. 2, No. 6, 2014, pp. 338-344. doi: 10.11648/j.ajls.20140206.12
References
[1]
Alessandro Riffel, Adriano Brandelli (2003). De-hairing activity of extracellular proteases produced by keratinolytic bacteria. J. of Chem. Technol. & Biotech. 78 (8), 855-859
[2]
Burtt, E. H., & Ichida, J. M.(1999).Bacteria useful for degrading keratin.US Patent 6214676.
[3]
Cantera, C. S., A. R. Angelinetti, G. Altobelli, and G. Gaita (1996). Hairsaving enzyme assisted dehairing. Influence of enzymatic products upon final leather quality. J. Soc. Leather Technol. Chem. 80:83–86.
[4]
Cappuccino, J.G. and Sherman, N. (2001). Microbiology: A Laboratory Manual. 6th Edition. Benjamin Cumings.CA.
[5]
Cassano, A., E. Drioli, R. Molinari, D. Grimaldiand M. Rossi (2000). Enzymatic membrane reactor for eco-friendly goat skin dehairing. J. Soc. Leat.Technol. Chem. 84:205–211.
[6]
Dhar S.C., Sreenivasulu S. (1984). Studies on the use of dehairing enzyme for ıts suitibilty in the preparation of improved animal feed. Leather Sci., 31: 261-267.
[7]
El-Refai, M.H.; Fatah,A.F.A. (2005). Improvement of the newly isolated Bacillus pumilus FH9 Keratinolytic activity. Process Biochem., 40, 2325-2332.
[8]
Evans KL, Crowder J, Miller ES (2000). Subtilisins of Bacillus spp. hydrolyze keratin And allow growth on feathers. Can. J. Microbiol. 6:1004-1011.
[9]
George, S., V. Raju, M. R. V. Krishnan, T. V. Subramanian, and K. Jayaraman(1995). Production of protease by Bacillus amyloliquefaciens in solid-state fermentation and its application in the dehairing of hides and skins. Process Biochem. 30:457–462.
[10]
Gessesse, A., Rajni, H.K., Gashe, B.A. (2003). Novel alkaline proteases from alkaliphilic bacteria grown on chicken feather. Enzyme Microb. Technol., 32 (5):519-524.
[11]
Hartree E.E. (1972). This modification makes the assay linear over a largerrange than the original assay. Anal. Biochem. 48:422
[12]
Ichida J. M., L. Krizova, C. A. LeFevre, H. M. Keener, D. L. Elwell & E. H. Burtt (2001). Journal of Microbiology Methods, 47, 199
[13]
Kim J.D. (2004). Purification and Characterization of a Keratinase from a Feather-Degrading Fungus, Aspergillus flavus Strain K-03. Microbiology, No. 35 (4), 219-225.
[14]
Lowry, O.H., Rosebrough, N.J., Farr, A.L., and Randall, R.J. (1951) J.Biol.Chem 193: 265
[15]
Madern, D. and Zaccai, G(2000).Halophilic adaptation of enzymes. Extremophiles, 4,91-98.
[16]
Mitsuiki, S., Sakai, M., Moriyama, Y., Goto, M., Furukawa, K. (2002). Purification and some properties of akeratinolytic enzyme from an alkaliphilic Nocardiopsis sp. TOA-1. Biosci. Biotechnol. Biochem. 66 (1), 164–167.
[17]
Mukhopadyay R.P., Chandra A.L. (1993). Protease of keratinolytic Streptomycetes to unhair goat skin. Indian J. Exp. Biol., 31: 557-558.
[18]
Nam and Y. R. Pyun(2002). Native-feather degradation by Fervidobacterium islandicum, a newly isolated keratinase-producing thermophilic anaerobe. Arch. microbiol. 178:538–547.
[19]
Purushotham, H., S. Malathi, P. V. Raoand K. V. Raghavan (1994). Dehairing enzyme by solid state fermentation. J. Soc. Leather Technol. Chem. 80:52–56.
[20]
Puvanakrishnan, R. and Dhar, S. C., (1986). Leather Sci., 33, 177–191
[21]
Raju AA, Chandrababu and Rao NM. (1996). Eco-friendly enzymatic dehairing using extracellular proteases from a Bacillus species. J Am Leather Chem. Assoc. 91: 115-119
[22]
[Rozs M, Manczinger L, Vágvölgyi C, Kevei F. (2001). Secretion of a trypsin- like thiol protease by a new keratinolytic strain of Bacillus licheniformis. 205: 221-224.
[23]
[Rozs M, Manczinger L, Vágvölgyi C, Kevei F. (2001). Secretion of a trypsin- like thiol protease by a new keratinolytic strain of Bacillus licheniformis. 205: 221-224.
[24]
Scopes, R.K., 1982. “Protein Purification: Principle and practice,” Springer-Verlag, New York, pp 195-196.
[25]
Sneath, P.H.A., Mair, N.S., Sharpe, M.E. & Holt, J.G., (1986). Bergey’s Manual of Systematic Bacteriology, Vol. 2 Baltimore: Williams and Wilkins. ISBN 0-683-07893-3
[26]
Suntornsuk W and Oda K (2005).Purification and characterization of keratinase from a thermotolerant feather- degrading bacterium. J. Ind. Microbiol. Biotechnol. 21: 1111-1117
[27]
THYS and BRANDELLI, A. (2004). Characterization of a protease of a feather-degrading. Microbacterium species. Letters in Applied Microbiology, February, 39 (2) 181-186.
[28]
Zerdani I, Faid M, Malki A. (2004). Feather wastes digestion by new isolated strains Bacillus sp. in Morocco. Afr. J. Biotech. 3: 67-70.
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