Alkaline protease production using eggshells and membrane-based substrates: Process modeling, optimization, and evaluation of detergent potency
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Abstract
This study focused on the use and eggshells and associated membranes (ESM) as a substrate for producing alkaline protease (AlkP). Furthermore, the study was performed to optimize the production process and check the feasibility of using the enzyme as a detergent. Bacillus mojavensis was used as a biological agent for fermentative enzyme production. The process was optimized with four factors at three levels using response surface methodology (RSM) coupled with the Box–Behnken design. Through RSM, optimal parameters for the production of AlkP were found to be the following: pH 9.08, temperature 39.74 °C, and ESM 197.8 g/l or 19.78% (w/v) with an incubation time of 48 h. Thereafter, the obtained enzyme was partially characterized. It retained more than 93.97% of its original activity in the pH range between 8 and 10, with optimal activity at pH 10. The minimum residual activity of AlkP was 39.6% of its original activity at pH 6.0. AlkP was found to be active at temperatures between 30 and 80 °C, but maximal at 60 °C. Residual activities were found to be 67.4%, 71.7%, 75.7%, 78.7%, and 78.7% in solutions with NaCl concentrations of 1.0, 1.5, 2, 2.5, and 3.0 M, respectively, at pH 8.0. A detergent compatibility study revealed that the obtained AlkP is quite effective as a detergent.
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References
Furhan J, Sharma S. Microbial alkaline proteases : findings and applications. Int J Invent Pharm Sci. 2014;2(4):823-34.
Singh R, Mittal A, Kumar M, Mehta PK. Microbial proteases in commercial applications. J Pharm Chem Biol Sci. 2016;4(3):365-74.
Haile G, Gessesse A. Properties of alkaline protease C45 produced by alkaliphilic Bacillus Sp. isolated from Chitu, Ethiopian Soda Lake. J Biotechnol Biomater. 2012;2(4):136.
Haile G. Alkaline proteaseproduction by an alkaliphilic bacterial isolate under solid state fermentation [thesis]. Addis Ababa: Addis Ababa University; 2009.
Hailemichael A, Gebremedhin B, Gizaw S, Tegegne A. Analysis of village poultry value chain in Ethiopia: implications for action research and development. Lives Working Paper 10. Nairobi: International Livestock Research Institute (ILRI); 2016.
Nilegaonkar SS, Zambare VP, Kanekar PP, Dhakephalkar PK, Sarnaik SS. Production and partial characterization of dehairing protease from Bacillus cereus MCM B-326. Bioresour Technol. 2007:(98):1238-45.
Nakano T, Ikawa N, Ozimek L. Chemical composition of chicken eggshell and shell membranes. Poult Sci. 2003;82(3):510-4.
King`ori AM. A review of the uses of poultry eggshells and shell membranes. Int J Poult Sci. 2011;10(11):908-12.
FAO. Poultry sector Ethiopia. Rome: FAO Animal Production and Health Livestock Country Reviews; 2019.
CSA. Report on small scale manufacturing industries survey. Addis Ababa: The Federal Democratic Republic of Ethiopia Central Statistical Agency; 2018.
Nagamalli H, Sitaraman M, Kandalai KK, Mudhole GR. Chicken egg shell as a potential substrate for production of alkaline protease by Bacillus altitudinis GVC11 and its applications. 3 Biotech. 2017;7(3):1-6.
Olajuyigbe FM. Optimized production and properties of thermostable alkaline protease from Bacillus subtilis SHS-04 grown on groundnut (Arachis hypogaea) meal. Adv Enzym Res. 2013;01(04):112-20.
Sarker PK, Talukdar SA, Deb P, Sayem SA, Mohsina K. Optimization and partial characterization of culture conditions for the production of alkaline protease from Bacillus licheniformis P003. Springerplus. 2013;2(1):506.
Marathe SK, Vashistht MA, Prashanth A, Parveen N, Chakraborty S, Nair SS. Isolation, partial purification, biochemical characterization and detergent compatibility of alkaline protease produced by Bacillus subtilis, Alcaligenes faecalis and Pseudomonas aeruginosa obtained from sea water samples. J Genet Eng Biotechnol. 2018;16(1):39-46.
Horikoshi K. Production of alkaline enzymes by alkalophilic microorganisms part ii alkaline amylase produced by bacillus no. A-40-2. Agric Biol Chem. 1971;35(11):1783-91.
Gessesse A, Gashe BA. Production of alkaline xylanase by an alkaliphilic Bacillus sp. isolated from an alkaline soda lake. J Appl Microbiol. 1997;83(4):402-6.
Gessesse A, Hatti-Kaul R, Gashe BA, Mattiasson B. Novel alkaline proteases from alkaliphilic bacteria grown on chicken feather. Enzyme Microb Technol. 2003;32(5): 519-24.
Warth AD, Scientific C, Ryde N. Relationship between the heat resistance of spores and the optimum and maximum growth temperatures of Bacillus Species. J Bacteriol. 1978; 134(3):699-705.
Devi RV, Jayaraman G, Rameshpathy M, Sridharan TB. Production and characterization of extracellular protease from halotolerant bacterium Virgibacillus dokdonesis Vitp14. Res J Biotechnol. 2012;7(2):38-42.
El-Hassayeb HEA, Abdel Aziz SMZ. Screening, production and industrial application of protease enzyme from marine bacteria. Int J Curr Microbiol Appl Sci. 2016;5(7):863-74.
Rai SK, Roy JK, Mukherjee AK. Characterisation of a detergent-stable alkaline protease from a novel thermophilic strain Paenibacillus tezpurensis sp. nov. AS-S24-II. Appl Microbiol Biotechnol. 2010;85(5):1437-50.
Hariharan M, Varghese N, Cherian AB, Sreenivasan PV, Paul J, Asmy Antony KA. Synthesis and characterisation of CaCO3 (Calcite) nano particles from cockle shells using chitosan as precursor. Int J Sci Res Publ. 2014;4(10):1-5.
Venkatesa PS, Gonfa G, Gizachew AK, Beyan SM, Ramesh G. Biosolubilization of Cr (VI) from tannery
sludge: process modeling, optimization, rate kinetics and thermodynamics aspects. Int J Recent Technol Eng. 2019;8(4):4808-16.
Bushell M. Manual of industrial microbiology and biotechnology. Enzyme Microb Technol. 1987;9(5):317.
Waites MJ, Morgan NL, Rockey JS, Higton G. Industrial microbiology: an introduction. Oxford: Blackwell Science; 2001.
Kumar CG, Tiwari MP, Jany KD. Novel alkaline serine proteases from alkalophilic Bacillus spp.: purification and some properties. Process Biochem. 1999;34(5):441-9.
Vadlamani S, Parcha SR. Studies on industrially important alkaline protease production from locally isolated superior microbial strain from soil microorganisms. Int J Biotechnol Appl. 2011;3(3):101-5.
Ramnani P, Suresh Kumar S, Gupta R. Concomitant production and downstream processing of alkaline protease and biosurfactant from Bacillus licheniformis RG1: bioformulation as detergent additive. Process Biochem. 2005;40(10):3352-9.
Kiranmayee R, Narasu LM. Alkaline protease from Bacillus firmus 7728. African J Biotechnol. 2007;6(21):2493-6.
Gupta R, Beg QK, Lorenz P. Bacterial alkaline proteases: molecular approaches and industrial applications. Appl Microbiol Biotechnol. 2002;59(1):15-32.
Abusham RA, Rahman R, Salleh A, Basri M. Optimization of physical factors affecting the production of thermo-stable organic solvent-tolerant protease from a newly isolated halo tolerant Bacillus subtilis strain Rand. Microb Cell Fact. 2009;8(1):20.
Biosci IJ, Ahmed M, Rehman R, Siddique A, Hasan F, Ali N. Production, purification and characterization of detergent-stable, halotolerant alkaline protease for eco-friendly application in detergents’ industry. Int J Biosci. 2016;8(2):47-65.
Haddar A, Sellami-Kamoun A, Fakhfakh-Zouari N, Hmidet N, Nasri M. Characterization of detergent stable and feather degrading serine proteases from Bacillus mojavensis A21. Biochem Eng J. 2010;51(2):53-63.