Application of modified silica gel in the process of trypsin immobilization
Justyna Miłek
Sylwia Kwiatkowska-Marks
Ilona Trawczyńska
Abstract
The paper presents the use of modified silica gel for the production of immobilized trypsin from bovine pancreas. Silica gel was modified with 3-aminopropyltriethoxysilane, followed by glutaraldehyde. The influence of stirring time on activity of the prepared biocatalyst was determined for individual stages of the modification. Activity of both native and immobilized trypsin was measured using Kunitz method. At the temperature of 55℃ and pH 7.6 native and immobilized trypsin onto modified silica gel indicate optimum activity. The influence of multiple recycling and storage time on activity of immobilized trypsin was tested. After fourteen days of storage at the temperature of 4℃ immobilized trypsin exhibits 75% of its initial activity.
Keywords:
trypsin, immobilization, silica gel, biocatalystReferences
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Methods of Enzymatic Analysis. 2012. Volume 2. Ed. H.-U. Bergmeyer. Elsevier, Science, Academic Press, p. 1020.
Caramori S.S., de Faria F.N., Viana M.P., Fernandes K.F., Carvalho L.B. Jr. 2011. Trypsin immobilization on discs of polyvinyl alcohol glutaraldehyde/polyaniline composite. Materials Science and Engineering, C, 31: 252–257.
Gomez J., Romero M. 2009. Adsorption of trypsin on commercial silica gel. Life Sciences, 9(4): 336-341.
Mohamad N.R., Marzuki N.H.C., Buang N.A., Huyop F., Waha R.A. 2015. Review. Agriculture and environmental biotechnology. An overview of technologies for immobilization of enzymes and surface analysis techniques for immobilized enzymes. Biotechnology and Biotechnological Equipment, 29(2): 205-220.
Monteiro F.M.F., Silva G.M.M. 2007. Immobilization of trypsin on polysaccharide film from Anacardium occidentale L. and its application as cutaneous dressing. Process Biochemistry, 42: 884-888.
Salar S., Mehrnejad F., Sajedi R.H., Arough J.M. 2017. Chitosan nanoparticles-trypsin interactions: Bio-physicochemical and molecular dynamics simulation studies. International Journal of Biological Macromolecules, 103: 902–909.
Shen S.-C., Ng W.K., Chia L., Dong Y.-C., Tan R.B.H. 2011. Sonochemical synthesis of (3-aminopropyl) triethoxysilane-modified monodispersed silica nanoparticles for protein immobilization. Materials Research Bulletin, 46: 1665–1669.
Sun X., Cai X., Wang R.-Q., Xiao J. 2015. Immobilized trypsin on hydrophobic cellulose decorated nanoparticles shows good stability and reusability for protein digestion. Analytical Biochemistry, 477: 21–27.
Yang G., Wu J., Xu G., Yang L. 2010. Comparative study of properties of immobilized lipase onto glutaraldehyde-activated amino-silica gel via different methods. Colloids and Surfaces B: Biointerfaces, 78: 351–356.
Zhou C., Wu X., Jiang B., Shen S. 2011. Immobilization strategy of accessible transmission for trypsin to catalyze synthesis of dipeptide in mesoporous suport. Korean Journal of Chemical Engineering, 28(12): 2300-2305.
Zhao Q., Hou Y., Gong G.-H., Yu M.-A., Jiang L., Liao F. 2010. Characterization of alcohol dehydrogenase from permeabilized brewer’s yeast cells immobilized on the derived attapulgite nanofibers. Applied Biochemistry and Biotechnology, 160(8): 2287–2299.
Miłek, J., Kwiatkowska-Marks, S., & Trawczyńska, I. (2019). Application of modified silica gel in the process of trypsin immobilization. Technical Sciences, 22(1), 35–43. https://doi.org/10.31648/ts.4346
Justyna Miłek
Sylwia Kwiatkowska-Marks
Ilona Trawczyńska
