PHYSICAL OPTIMIZATION OF THERMOSTABLE ALKALINE PROTEASE BY E. COLI BL21 (DE3) PLYSS HARBORING 50A PROTEASE GENE USING RESPONSE SURFACE METHODOLOGY
DOI:
https://doi.org/10.11113/jt.v78.4048Keywords:
Optimization, Response Surface Methodology, thermostable proteaseAbstract
Physical optimization is important for enzyme production by fermentation process. In general, fermentation process at optimal condition increases the expression and production level of enzyme to many times in comparison with their natural production. This study was focused on the optimization of the physical factors that influenced the thermostable alkaline protease production. The induction and incubation time were studied using conventional method while the other three factors which are incubation temperature, initial pH of medium and agitation speed were optimized by response surface methodology (RSM). The interaction effects among these factors were explained using response plot and the model adequacy was satisfactory as the coefficient of determination (R2) was 96.48%. The enhancement of thermostable protease from 197.83 U/ml to 325.89 U/ml was achieved using both conventional and statistical approach of response surface methodology (RSM). This present study proved that physical optimization significantly affects the protease production and the optimum physical condition obtained may applied in large scale process.
References
Esposito, T. S., Amiral, I. P. G., Buarque, D. S., Oliveira, G. B., Carvalho Jr, L. B. and Bezerra, R. S. 2009. Fish Processing Waste as a Source of Alkaline Proteases for Laundry Detergent. Food Chemistry. 112: 125-130.
Abidi, F., Chobert, J. M., Haertle, T. and Marzouki, M. N. 2011. Purification and Biochemical Characterization of Stable Alkaline Protease Prot-2 from Botrytis cinerea. Process Biochemistry. 46: 2301-2310.
Jain, D., Pancha, I., Mishra, S. K., Shrivastav, A. and Mishra, S. 2012. Purification and Characterization of Haloalkaline Thermoactive, Solvent Stable and SDS-Induced Protease from Bacillus Sp: A Potential Additive for Laundry Detergents. Bioresource Technology. 115: 228-236.
Gupta, R., Beg, Q. K. and Lorenz, P. 2002. Bacterial Alkaline Proteases: Molecular Approaches and Industrial Applications. Applied Microbiology Biotechnology. 59: 15-32.
Saeki, K., Ozaki, K., Kobayashi, T. and Ito, S. 2007. Review: Detergent Alkaline Protease: Enzymatic Properties, Genes and Crystal Structures. Journal of Bioscience and Bioengineering. 103(6): 501-508.
Akcan, N. 2012. Production of Extracellular Protease in Submerged Fermentation by Bacillus licheniformis ATCC 12759. African Journal of Biotechnology. 11(7): 1729-1735.
Liu, S., Fang, Y., Lv, M., Wang, S. and Chen, L. 2010. Optimization of the Production of Organic Solvent-Stable Protease by Bacillus sphaericus DS11 with Response Surface Methodology. Bioresource Technology. 101: 7924-7929.
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.
Mannan, S., Fakhru’l-Razi, A. & Alam, M. Z. 2007. Optimization of Process Parameters for the Bioconversion of Activated Sludge by Penicillum corylophilum, using Response Surface Methodology. Journal of Environmental Science. 19: 23-28.
Ibrahim, N. A and Yusoff, N., 2013. Thermostable Alkaline Serine Protease from Thermophilic Bacillus species. International Research Journal of Biological Sciences. 2(2): 29-33.
Razak, C., Rahman, R., Salleh, A. B., Yunus, W., Ampon, K. and Basri, M. 1995. Production of a Thermostable Protease from a New High Ph Isolate of B. stearothermophilus. Journal of Biociences. 6: 94-100.
Fu, Z., Hamid, S. A., Razak, C. N. A., Basri, M., Salleh, A. B. and Rahman, R. N. Z. A. 2003. Secretory Expression in Escherichia Coli and Single-Step Purification of a Heat Stable Alkaline Protease. Protein Expression and Purification. 28: 63-68.
Vasquez-Bahena, J. M., Vega-Estrada, J., Santiago-Hernandez, J. A., Ortega-Lopez, J., Flores-Cotera, L. B., Montes-Horcasitas, M. C., et al. 2006. Expression and improved production of the soluble extracellular invertase from Zymomo nas mobilis in Escherichia coli. Enzyme and Microbial Technology. 40: 61-66.
Raj, A., Khess, N., Pujari, N., Bhattacharya, S., Das, A. and Rajan, S. S. 2012. Enhancement of Protease Production by Pseudomonas aeruginosa Isolated from Dairy Effluent Sludge and Determination of Its Fibrinolytic Potential. Asian Pacific Journal of Tropical Biomedicine. 1845-1851.
Thys, R. C. S., Guzzon, S. O., Cladera-Olivera, F. and Brandelli, A. 2006. Optimization of Protease Production by Microbacterium sp. in Feather Meal Using Response Surface Methodology. Process Biochemistry. 41: 67-73.
Bauman, R. W. 2007. Microbiology with Disease by Taxonomy. 2nd ed. San Francisco:Pearson Benjamin Cummings.
Loc, N. H., Quang, H. T., Lam, B. T. H. and Trang, D. T. T. 2011. The Effects of Culture Conditions on Neutral Protease (NPRC10) in a Recombinant Escherichia coli BL21(DE3). Scholar Research Library. 2(3): 474-485.
Romsomsa, N., Chim-anagae, P. and Jangchud, A., 2010. Optimization of Silk Degumming Protease Production from Bacillus subtilis C4 using Plackett-Burman Design and Response Surface Methodology. ScienceAsia. 36: 118-124.
Calik, P., Bilir, E., Calik, G. and Ozdamar, T. H. 2002. Influence of pH Conditions on Metabolic Regulations In Serine Alkaline Protease Production by Bacillus licheniformis. Enzyme and Microbial Technology. 31: 685-697.
Rahman, R. N. Z. A., Geok, L. P, Basri, M. and Salleh, A. B. 2005. Physical Factors Affecting the Production of Organic Solvent-Tolerant Protease by Pseudomonas aeruginosa strain K. Bioresource Technology. 96: 429-436.
Shafee, N., Aris, S. N., Rahman, R. N. Z. A., Basri, M. and Salleh, A. B. 2005. Optimization of Environmental and Nutritional Conditions for the Production of Alkaline Protease by a Newly Isolated Bacterium Bacillus cereus strain 146. Journal of Applied Sciences Research. 1(1): 1-8.
Beg, Q. K., Sahai, V. and Gupta, R. 2003. Statistical Media Optimization and Alkaline Protease Production from Bacillus mojavensis in a Bioreactor. Process Biochemistry. 39: 203-209.
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