Biochemical Characteristics of Immobilized Trypsin from Porgy (Stenotomus chrysops) Viscera
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Published:
Aug 31, 2020
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Abstract
Partially purified trypsin from porgy (Stenotomus chrysops) viscera was immobilized onto Octyl Sepharose CL-4B. The optimal temperature and pH for the hydrolysis of N-α-benzoyl-DL-arginine-p-nitroanilide (DL-BAPNA) by the immobilized trypsin were 55oC and 8, respectively. The immobilized trypsin exhibited a much broader pH range (6-11), temperature (20oC-80oC) activity and stability versus the free form of the enzyme. The immobilized enzyme was less sensitive to inhibition by the soybean trypsin inhibitor. According to the results, the immobilized trypsin and free enzyme retained 70.59% and 20.10% of their activity, respectively, when they were incubated with 1 µM of the soybean trypsin inhibitor. For the reusability study, the immobilized trypsin maintained 41.31% of its activity after 6 periods of activity, indicating that the immobilized trypsin had appropriate stability and could be reused.
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How to Cite
[1]
Poonsin, T., M. Barman, A. , K. Simpson, B. and Klomklao, S. 2020. Biochemical Characteristics of Immobilized Trypsin from Porgy (Stenotomus chrysops) Viscera. Journal for Community Development and Life Quality. 8, 3 (Aug. 2020), 530–543.
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Research Articles
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References
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Koide, T. and T. Ikenaka. 1973. Studies on soybean trypsin inhibitors: 1. Fragmentation of soybean trypsin inhibitor (Kunitz) by limited prateolysis and by chemical cleavage. European Journal of Biochemistry 32(3): 401-407.
Lage, F.A.P., J.J. Bassi, M.C. Corradini, L.M. Todero, J.H. Luiz and A.A. Mendes. 2016. Preparation of a biocatalyst via physical adsorption of lipase from Thermomyces lanuginosus on hydrophobic support to catalyze biolubricant synthesis by esterification reaction in a solvent-free system, Enzyme and Microbial Technology 84: 56-67.
Li D., N.A. Ackaah-Gyasi and B.K. Simpson. 2013. Immobilization of bovine trypsin onto controlled pore glass. Journal of Food Biochemistry 38(2): 184-195.
Purcena, L.L.A., S.S. Caramori, S. Mitidieri and K.F. Fernandes. 2009. The immobilization of trypsin onto polyaniline for protein digestion. Materials Science and Engineering: C 29(4): 1077-1081.
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Sears, P.S. and D.S. Clark. 1993. Comparison of soluble and immobilized trypsin kinetics: Implications for peptide synthesis. Biotechnology and Bioengineering 42(1): 118-124.
Shtelzer, S., S. Rappoport, D. Avnir, M. Ottolenghi and S. Braun. 1992. Properties of trypsin and of acid phosphatase immobilized in sol-gel glass matrices. Biotechnology and Applied Biochemistry 15(3): 227-235.
Simpson, B.K. 2000. Digestive proteinases from marine animals. pp. 531-540. In: N.F. Haard and B.K. Simpson (eds). Seafood Enzymes: Utilization and Influence on Postharvest Seafood Quality. Marcel Dekker, New York.
Steel, R.G.D. and J.H. Torrie. 1980. Principles and Procedures of Statistics: A Biometrical Approach. McGraw-Hill, New York.
Sun, X., Cai X., R.Q. Wang and J. Xiao. 2015. Immobilized trypsin on hydrophobic cellulose decorated nanoparticles shows good stability and reusability for protein digestion. Analytical Biochemistry 477: 21-27.
Vidinha, P., V. Augusto, M. Almeida, I. Fonseca, A. Fidalgo, L. Ilharco, J.M. Cabral and S. Barreiros. 2006. Sol-gel encapsulation: an efficient and versatile immobilization technique for cutinase in non-aqueous media. Journal of Biotechnology 121(1): 23-33.
Wu, N., S. Wang, Y. Yang, J. Song, P. Su and Y. Yang. 2018. DNA-directed trypsin immobilization on a polyamidoamine dendrimer-modified capillary to form a renewable immobilized enzyme microreactor. International Journal of Biological Macromolecules 113(1): 38-44.
Wu, S., B. Liu and S. Li. 2005. Behaviors of enzyme immobilization onto functional microspheres. International Journal of Biological Macromolecules 37(5): 263-267.
Atacan, K., B. Çakıroglu and M. Ozacar. 2017. Covalent immobilization of trypsin onto modified magnetite nanoparticles and its application for casein digestion. International Journal of Biological Macromolecules 97: 148-155.
Azevedo, R.D.S., I.P.G. Amaral, A.C.M. Ferreira, T.S. Espósito and R.S. Bezerra. 2018. Use of fish trypsin immobilized onto magnetic-chitosan composite as a new tool to detect antinutrients in aquafeeds. Food Chemistry 257: 302-309.
Bassan, J.C., T.M. de Souza Bezerra, G. Peixoto, C.Z.P. da Cruz, J.P.M. Galan, A.B.D.S. Vaz, S.S. Garrido, M. Filice and R. Monti. 2016. Immobilization of trypsin in lignocellulosic waste material to produce peptides with bioactive potential from whey protein. Materials 9(5): 357. Doi: 10.3390/ma9050357.
Bayramoglu, G., M. Yılmaz, A.U. Şenel and M.Y. Arıca. 2008. Preparation of nanofibrous polymer grafted magnetic poly(GMA-MMA)-g-MAA beads for immobilization of trypsin via adsorption. Biochemical Engineering Journal 40(2): 262-274.
Bougatef, A. 2013. Trypsins from fish processing waste: characteristics and biotechnological applications-comprehensive review. Journal of Cleaner Production 57: 257-265.
Erlanger, B.F., N. Kokowsky and W. Cohen. 1961. The preparation and properties of two new chromogenic substrates of trypsin. Archives of Biochemistry and Biophysics 95(2): 271-278.
Ganachaud, C., D. Bernin, D. Isaksson and K. Holmberg. 2013. An anomalous behavior of trypsin immobilized in alginate network. Applied Microbiology and Biotechnology 97(10): 4403-4414.
Garcia-Galan, C., A. Berenguer-Murcia, R. Fernandez-Lafuente and R.C. Rodrigues. 2011. Potential of different enzyme immobilization strategies to improve enzyme performance. Advanced Synthesis and Catalysis 353(16): 2885-2904.
Kang, K., C. Kan, A. Yeung and D. Liu. 2005. The properties of covalently immobilized trypsin on soap-free P(MMA-EA-AA) latex particles. Macromolecular Bioscience 5(4): 344–351.
Klomklao, S., S. Benjakul, W. Visessanguan, H. Kishimura and B.K. Simpson. 2007. Trypsin from pyloric caeca of bluefish (Pomatomus saltatrix). Comparative Biochemistry and Physiology B 148(4): 382-389.
Koide, T. and T. Ikenaka. 1973. Studies on soybean trypsin inhibitors: 1. Fragmentation of soybean trypsin inhibitor (Kunitz) by limited prateolysis and by chemical cleavage. European Journal of Biochemistry 32(3): 401-407.
Lage, F.A.P., J.J. Bassi, M.C. Corradini, L.M. Todero, J.H. Luiz and A.A. Mendes. 2016. Preparation of a biocatalyst via physical adsorption of lipase from Thermomyces lanuginosus on hydrophobic support to catalyze biolubricant synthesis by esterification reaction in a solvent-free system, Enzyme and Microbial Technology 84: 56-67.
Li D., N.A. Ackaah-Gyasi and B.K. Simpson. 2013. Immobilization of bovine trypsin onto controlled pore glass. Journal of Food Biochemistry 38(2): 184-195.
Purcena, L.L.A., S.S. Caramori, S. Mitidieri and K.F. Fernandes. 2009. The immobilization of trypsin onto polyaniline for protein digestion. Materials Science and Engineering: C 29(4): 1077-1081.
Seabra, I.J. and M.H. Gil. 2007. Cotton gauze bandage: A support for protease immobilization for use in biomedical applications. Revista Brasileira de Ciências Farmacêuticas 43(4): 535-542.
Sears, P.S. and D.S. Clark. 1993. Comparison of soluble and immobilized trypsin kinetics: Implications for peptide synthesis. Biotechnology and Bioengineering 42(1): 118-124.
Shtelzer, S., S. Rappoport, D. Avnir, M. Ottolenghi and S. Braun. 1992. Properties of trypsin and of acid phosphatase immobilized in sol-gel glass matrices. Biotechnology and Applied Biochemistry 15(3): 227-235.
Simpson, B.K. 2000. Digestive proteinases from marine animals. pp. 531-540. In: N.F. Haard and B.K. Simpson (eds). Seafood Enzymes: Utilization and Influence on Postharvest Seafood Quality. Marcel Dekker, New York.
Steel, R.G.D. and J.H. Torrie. 1980. Principles and Procedures of Statistics: A Biometrical Approach. McGraw-Hill, New York.
Sun, X., Cai X., R.Q. Wang and J. Xiao. 2015. Immobilized trypsin on hydrophobic cellulose decorated nanoparticles shows good stability and reusability for protein digestion. Analytical Biochemistry 477: 21-27.
Vidinha, P., V. Augusto, M. Almeida, I. Fonseca, A. Fidalgo, L. Ilharco, J.M. Cabral and S. Barreiros. 2006. Sol-gel encapsulation: an efficient and versatile immobilization technique for cutinase in non-aqueous media. Journal of Biotechnology 121(1): 23-33.
Wu, N., S. Wang, Y. Yang, J. Song, P. Su and Y. Yang. 2018. DNA-directed trypsin immobilization on a polyamidoamine dendrimer-modified capillary to form a renewable immobilized enzyme microreactor. International Journal of Biological Macromolecules 113(1): 38-44.
Wu, S., B. Liu and S. Li. 2005. Behaviors of enzyme immobilization onto functional microspheres. International Journal of Biological Macromolecules 37(5): 263-267.
