Statistical Optimization of the production of protease from Bacillus sp. MSK-01 in submerged fermentation
DOI:
https://doi.org/10.5912/jcb810Keywords:
Bacillus sp. MSK-01, Protease, Submerged fermentation, Response surface methodologyAbstract
Protease has been in increasing demand in industries due to its hydrolytic nature. In industries, high yield of enzyme is required to meet the industrial need at a relatively cheaper cost. In the present study, the protease from Bacillus sp. MSK-01 was produced in large quantity by submerged fermentation. Statistical techniques including Plackett-Burman and Response surface methodology are useful tools for optimizing many parameters at a time and are used for increasing the protease production from Bacillus sp. MSK-01. 19 different parameters were chosen, out of which 15 factors had positive effect on protease yield. Four maximum influencing factors were peptone, magnesium sulphate, skim milk powder and casein were chosen to further increase the protease yield. 397.3 IU ml-1 of enzyme yield was obtained under optimized conditions which lead to 198 fold increase in the yield of protease from unoptimized condition.
References
Hartley, BS. (1960) Proteolytic enzymes. Annual Review in Biochemistry 29: 45–72.
Nasuno, S. and Ohara, T. (1971) Hyperproduction of proteinase and some hydrolytic enzymes by mutants of Aspergillus sojae. Agricultural and Biological Chemistry 35: 829-835.
Malathi, S. and Chakraborty, R. (1991) Production of alkaline protease by a new Aspergillus flavus isolate un-der solid substrate fermentation conditions for use as a depilation agent. Applied and Environmental Microbiology 57: 712-716.
Monod, M., Togni, G., Rahalison, L., Frenk, E. (1991) Isolation and characterization of an extracellular alkaline protease of Aspergillus fumigatus. Journal of Medical Microbiology 35: 23-28.
Luisetti, M., Piccioni, PO., Dyne, K., Donnini, M., Bulgheroni, A., Pasturenzi, L., Donnetta, AM. and Peona, V. (1991) Some properties of the alkaline proteinase from Aspergillus melleus. International Journal of Tissue Reaction 13: 187-192.
Barthomeuf, C., Pourrat, H. and Pourrat, A. (1992) Col-lagenolytic activity of a new semi-alkaline protease from Aspergillus niger. Journal of Fermentation and Bioengineering 73: 233-236.
Larcher, G., Bouchara, JP., Annaix, V., Symoens, F., Chabasse, D. and Tronchin, G. (1992) Purification and characterization of a fibrinogenolytic serine proteinase from Aspergillus fumigatus culture filtrate. FEBS Letters 308: 65-69.
Godfrey, T. and West, S. (1996) Introduction to industrial enzymology. Industrial enzymology, Mac Millan Press, London, 1-8.
Dubey, R., Adhikary, S., Kumar, J. and Sinha, N. (2010) Isolation, Production, Purification, Assay and Characterization of Alkaline Protease Enzyme from Aspergillus niger and its Compatibility with Commercial Detergents. Developmental Microbiology and Molecular Biology 1: 75-94.
Rao, M.B., Tanksale, A.M., Ghatge, M.S. and Deshpande, V.V. (1998) Molecular and biotechnological aspects of microbial proteases. Microbiology and Molecular Biology Reviews 62: 597-635.
Gupta, R., Beg, Q.K., Lorenz, P. (2002) Bacterial alkaline proteases: molecular approaches and industrial applications. Applied Microbiology and Biotechnology 59: 15–32.
Sabotic, J. and Kos, J. (2012) Microbial and fungal protease inhibitors current and potential applications. Applied Microbiology and Biotechnology 93:1351-1375.
Kumar, C.G. and Takagi, H. (1999) Microbial alkaline proteases from a bioindustrial view point. Biotechnology Advance 17: 561-594.
Jellouli, K., Bougatef, A., Manni, L., Agrebi, R., Siala, R. and Younes, I. (2009) Molecular and biochemical characterization of an extracellular serine-protease from Vibrio metschnikovii J1. Journal of Industrial Microbiology & Biotechnology 36(7): 939–948.
Atalo, K. and Gashe, B.A. (1993). Protease production by a thermophilic Bacillus sp. (P-001A) which degrades various kinds of fibrous proteins. Biotechnology Letters 11: 1151-1156.
Lowry, O.H., Rosebrough, N., Farr, A.L. and Rondall, R.L. (1951) Protein measurement with the Folin phenol reagent. Journal of Biological Chemistry 193: 265–273.
Yadav, S.K., Bisht, D., Singh, R., Gaur, R. and Darmwal, N.S. (2013) Development of bioprocess for improved production of alkaline protease by mutant strain of Aspergilllus flavus in solid state fermentation using agricultural wastes. The Internet Journal of Microbiology 12(1).
Oskouie, S.F.G., Tabandeh, F., Yakhchali, B. and Eftekhar, F. (2008) Response surface optimization of medium composition for alkaline protease production by Bacillus clausii. Biochemical Engineering Journal 39: 37-42.
Bas, D. and Boyaci, I.H. (2007) Modelling and optimization I: Usability of Response Surface Methodology. Journal of Food Engineering 74: 836-845.
Geiger, E.O., Vogel, H.C. and Todaro, C.L. (1996) In: Fermentation and Biochemical Engineering Handbook; Ed. Noyes Publications, USA, 161-180.
Carvalho, C.M.L., Serralheiro, M.L.M., Cabral, J.M.S. and Airebarros, M.R. (1997) Application of factorial design to the study of trans esterification reactions using cutinase in AOT reversed micelles. Enzyme and Microbial Technology 21: 117–123.
Li, J., Ma, C., Ma, Y., Li, Y., Zhou, W. and Xu, P. (2007) Medium optimization by combination of response surface methodology and desirability function: an application in glutamine production. Applied Microbiology and Biotechnology 74: 563–571.
Cheng, S.W., Wang, Y.F. and Wanga, M.L. (2012) Statistical optimization of medium compositions for alkaline protease production by newly isolated Bacillus amyloliquefaciens. Chemical and Biochemical Engineering 26(3): 225–231.
Akolkar, A., Bharambe, N., Trivedi, S. and Desai, A. (2009) Statistical optimization of medium components for extracellular protease production by an extreme haloarchaeon, Alobacterium sp. SP1 (1). Letters in Applied Microbiology 48(1):77-83.
Reddy, L.V.A., Wee, Y.J., Yun, J.S. and Ryu, H.W. (2008) Optimization of alkaline protease production by batch culture of Bacillus sp. RKY3 through Plackett–Burman and response surface methodological approaches. Bioresource Technology 99: 2242–2249.
Moorthy, I.M. and Bhaskar, R. (2013) Statistical modeling and optimization of alkaline protease production from a newly isolated alkalophilic Bacillus sp. BGS using response surface methodology and genetic algorithm. Preparative Biochemistry and Biotechnology 43(3): 293-314.
Kumar, P.K.P., Mathivanan, V., Karunakaran, M., Renganathan, S. and Sreenivasan, R.S. (2008) Studies on the effects of pH and incubation period on protease production by Bacillus sp. using groundnut cake and wheat bran. Indian Journal of Science and Technology 1 (4): 1-4.
Saxena, R.K., Dutt, K., Meghwanshi, G.K. and Gupta, P. (2008) Role of casein on induction and enhancement of production of a bacterial milk clotting protease from an indigenously isolated Bacillus subtilis. Letters in applied Microbiology Vol 46(5): 513-518.
