Enhancement of protease production by Bacillus sp. and Micrococcus varians induced by UV-mutagenesis

Enhancement of protease production by Bacillus sp. and Micrococcus varians induced by UV-mutagenesis ( Vol-2,Issue-5,September - October 2017 )

Author: Chibani Hiba Rahman, Fellahi Soltana, Chibani Abdelwaheb

ijeab doi crossref DOI: 10.22161/ijeab/2.5.10

Keyword: Bcillus sp., Micrococcus varians,protease, UV-mutagenesis.

Abstract: Microbial proteases contribute nearly 40% of the total worldwide enzyme market. Hence, with the view of this significance, the main objective of the present study was to enhance protease production of two bacterial strains, Bcillus sp. and Micrococcus varians using UV mutagenesis. Induction of mutation in both strains was carried out at different exposure times: 0, 3, 6, 9, 12, 15, 18 and 21 min at a distance of 10 between UV source and treated bacteria.Two best protease producer mutants for the two bacterial strains (UV-9 for Bcillus sp.and UV-18 forMicrococcus varians) were selected based on the clearance zone diameter of mutant colonies on 1% skimmed milk agar plates. UV-9 mutant showed 1.4 fold higher protease activity than the wild type in solid and liquid medium. However UV-18 mutant was found to produce 2.5 fold increases over the wild type on agar plates and 2.1 fold enhancement in liquid-medium assay.The two mutants were very effective in feather keratin-degrading in less than two days, UV- 18 was more efficient than UV-9.


[1] Bernhard, K., Schrempf, H., and Goebel, W. 1978.Bacteriocin and antibiotic resistance plasmids in Bacillus cereus and Bacillus subtilis.J.bacteriol.,133(2), 897-903.
[2] Boominadhan, U., Rajakumar; R., Sivakumar, P.K.V. and Melvin, M.J. 2009. Optimization of protease enzyme production using Bacillus sp. isolated from different wastes.Bot Res Intl., 2: 83-87.
[3] Chellappan, S., Jasmin, C., Basheer, S.M., Elyas, K.K., Bhat, S.G., et al. 2006. Production, purification and partial characterization of a novel protease from marine Engyodontium album BTMFS10 under solid state fermentation. ProcBiochem., 41: 956-961.
[4] Hopwood, D.A., Bibb, M.J.,Chater,K.F.,Kieser, T.,Bruton,C.J., Kieser,H.M., Lydiate,D.J., Smith,C.P., Ward,J.M. and Schrempf,H. 1985. Genetic manipulation of Streptomyces: a laboratory manual, John Innes Foundation Norwich,pp, 356.
[5] Dutta, J. R.and Banerjee, R. 2006.Isolation and characterization of a newly isolated Pseudomonas mutant for protease production.Braz Arch Biol Technol., 49(1), 37-47.
[6] Glazer, A.N.andNikaido, H. 1995.Microbial enzymes. In: Glazer AN, Nikaido H (eds) Microbial Biotechnology, Freeman and Co,New York: W.H, pp. 24–263.
[7] Gupta, R., Beg, Q.K. And Lorenz, P.2002. Bacterial alkaline proteases: molecular approaches and industrial applications. Appl. Microbiol. Biotech., 59:15-32.
[8] Javed, S., Meraj, M., Bukhari, S.A., Irfan, R. andMahmood, S. 2013. Hyper-production of Alkaline Protease by Mutagenic Treatment of Bacillus subtilisM-9 using Agroindustrial Wastes in Submerged Fermentation. J MicrobBiochem Technol., 5: 074-080.
[9] Johnvesly, B., Manjunatha, B.R. and Naik, G.R. 2002.Pigeon pea waste as a novel, inexpensive substrate for production of thermostable alkaline protease from thermoalklophilicBacillus sp. JB-99.Bioresour Technol., 82, 61-64.
[10] Justin, C., Khodursky, A., Peter, B., Brown, P.O. and Hanawalt, P.C. 2001.Comparative gene expression profilesfollowing UV exposure in wild type and SOS-deficient Escherichia coli. Genetics., 158: 41-64.
[11] Karn, N.and Karn, S. K. 2014.Evaluation and Characterization of Protease Production by Bacillus sp. Induced By UV-Mutagenesis.Enz Eng.,3(119), 2.
[12] Nadeem, M., Qazi, J. I. and Baig, S. 2010. Enhanced production of alkaline protease by a mutant of Bacillus licheniformis N-2 for dehairing.Braz Arch Biol Technol., 53(5), 1015-1025.
[13] Rao, M.B., Tanksale, A.M., Ghatge, M.S. and Deshpande, V.V. 1998.Molecular and Biotechnological Aspects of Microbial Proteases.Microbiol. Mol. Biol. Rev., 62:597-635.
[14] Reddy, L.V., Wee, Y.J., Yun, J.S., Ryu, H.W. 2008. Optimization of alkaline protease production by batch culture of Bacillus sp. RKY3 through Plackett-Burman and response surface methodological approaches. Bioresour Technol., 99(7):2242-2249.
[15] Schallmey, M., Singh, A. and Ward, O.P. 2004.Developments in the use of Bacillus species for industrial production. Can. J.Microbiol., 50(1): 1-7.
[16] Shikha, S.A.and Darmwal, N.S. 2007.Improved production of alkaline protease from a mutant of alkalophilic Bacillus pantotheneticus using molasses as a substrate.Bioresour Technol., 98(4), 881-5.
[17] Solaiman, E.A.M., Hegazy, W.K. andMoharam, M.E. 2005.Induction of overproducing alkaline protease Bacillus mutants through UV irradiation. Arab J. Biotech.,8 (1): 49-60.
[18] Wang, S. L., Hsu, W. T., Liang, T. W., Yen, Y. H. and Wang, C. L. 2008.Purification and characterization of three novel keratinolyticmetalloproteases produced by ChryseobacteriumindologenesTKU014 in a shrimp shell powder medium.Bioresour Technol.,99(13), 5679-5686.
[19] Yang, S. and Huang, C. I. 1994.Protease production by amylolytic fungi in solid statefermentation. J. Chin. Agric. Chem. Soc., 32:589-601.

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