RELATIVE CHARACTERIZATION OF IMMOBILIZED BETA-GLUCOSIDASE ON CALCIUM-ALGINATE AND ACID FUNCTIONALIZED MULTIWALLED CARBON NANOTUBES
DOI:
https://doi.org/10.11113/jt.v81.13314Keywords:
β-glucosidase, MWCNT, Ca-alginate, immobilization, kineticsAbstract
Natural polymeric gel such as alginate and carrageenan, and carbon nanomaterials, especially MWCNT and graphene are gaining increasing popularity as immobilization support owing to their biocompatibility. Thus, this study attempts to make a relative characterization of the immobilized β-glucosidase on alginate and multiwalled carbon nanotubes (MWCNT) to assess their relative merits as biocatalyst. Acid functionalization of MWCNT lead to the formation of carboxyl groups at the MWCNT surface which seemed to have role in the stable attachment of the enzyme on the MWCNT surface. The effectiveness of the immobilized enzyme was evaluated using hydrolysis of p-nitrophenyl-β-D-glucopyranoside to p-nitrophenol. The optimum immobilization conditions were found to be 17 mg MWCNT and 4 h incubation time for MWCNT, and 3.5 wt% sodium alginate solution and 2 h incubation time for the Ca-alginate beads. The immobilization yield on MWCNT was found to be 96% and that for the Ca-alginate beads was 86%. The residual activity of the enzyme at the third cycle of subsequent reuse was 85% for the enzyme-MWCNT and almost 50% for the enzyme-alginate beads. Both, Ca-alginate and MWCNT immobilized enzyme were found to follow Michaelis-Menten kinetics. The kinetic data were best represented by the Langmuir plot. The kinetic constants Vmax and Km for the MWCNT immobilized enzyme were obtained as 1.324 mM/min and 0.801 mM, respectively, while for the alginate immobilized enzyme, the constants predicted were 1.240 mM/min and 0.524 mM, respectively. Thus MWCNT immobilized β-glucosidase exhibited greater stability in terms of activity and reusability compared to Ca-alginate beads.
References
Ansari, S. A., Sattar, R., Chibber, S., and Khan, M. J. 2013. Enhanced Stability of Luyveromyces β Galactosidase Immobilized on Glutaraldehyde Modified Multiwalled Carbon Nanotubes. Journal of Molecular Catalysis B: Enzymatic. 97: 258-263.
https://doi.org/10.1016/j.molcatb.2013.09.008
Busto, M. D., Ortega, N., and Perez-Mateos, M. 1997. Effect of Immobilization on the Stability of Bacterial and Fungal β-d-glucosidase. Process Biochemistry. 32(5): 441-449.
Tan, H., Feng, W., and Ji, P. 2012. Lipase Immobilized on Magnetic Multi-walled Carbon Nanotubes. Bioresource Technology. 115: 172-176.
https://doi.org/10.1016/S0032-9592(96)00104-5.
Tan, I. S., and Lee, K. T. 2015. Immobilization of β-glucosidase from Aspergillus Niger on κ-carrageenan Hybrid Matrix and its Application on the Production of Reducing Sugar from Macroalgae Cellulosic Residue. Bioresource Technology. 184: 386-394.
http://dx.doi.org/10.1016/j.biotech.2014.10.146.
Abdulla, R., and Ravindra, P. 2013. Characterization of Cross Linked Burkholderia Cepacia Lipase in Alginate and κ-carrageenan Hybrid Matrix. Journal of the Taiwan Institute of Chemical Engineers. 44: 545-55.http://dx.doi.org/10.1016/j.jtice.2013.01.003.
Jung, Y. R, Shin, H. Y., Song, Y. S., Kim, S. B., and Kim, S. W. 2012. Enhancement of Immobilized Enzyme Activity by Pretreatment of β-glucosidase with Cellobiose and Glucose. Journal of Industrial and Engineering Chemistry. 18: 702-706.
https://doi.org/10.1016/j.jiec.2011.11.133.
Tumturk, H., Karaca, N., Demirel, G., and Sahin, F. 2007. Preparation and Application of Poly(n,n-dimethylacrylamide-co-acrylamide) and Poly(n-isopropylacrylamide-co-acrylamide)/κ-carrageenan Hydrogels for Immobilization of Lipase. International Journal of Biological Macromolecules. 40: 281-285.
https://doi.org/10.1016/j.ijbiomac.2006.07.004.
Filiz Kara, F., Demirel, G., and Hayrettin, T. 2006. Immobilization of Urease by using Chitosan–alginate and Poly(acrylamide-co-acrylic acid)/κ-carrageenan Supports. Bioprocess and Biosystems Engineering. 29: 207-211.
Datta, S., Christena, L. R., and Rajaram, Y. R. S. 2013. Enzyme Immobilization: An Overview on Techniques and Support Materials. Biotech. 3: 1-9.
Feng, W. and Ji, P. 2011. Enzymes Immobilized on Carbon Nanotubes. Biotechnology Advances. 29: 889-895.
https://doi.org/10.1016/j.biotechadv.2011.07.007.
Sheldon, R. A. 2007. Enzyme Immobilization. Advanced Synthesis & Catalysis. 349: 1289-1307.
Li, L., Feng, W., and Pan, K. 2013. Immobilization of Lipase on Amino-cyclodextrin Functionalized Carbon Nanotubes for Enzymatic Catalysis at the Ionic Liquid-organic Solvent Interface. Colloids and Surfaces B: Biointerfaces. 102: 124-129. https://doi.org/10.1016/j.colsurfb.2012.07.050.
Mubarak, N. M., Wong, J. R., Tan, K. W., Sahu, J. N., Abdullah, E. C., Jayakumar, N. S., and Ganesan, P. 2014. Immobilization of Cellulose Enzyme on Functionalized Multiwall Carbon Nanotubes. Journal of Molecular Catalysis B: Enzymatic. 107: 124-131.
https://doi.org/10.1016/j.molcatb.2014.06.002.
Prlainovic, N. Z., Bezbradica, D. I., Rogan, J. R., Uskokovic, P. S., Mijin, D. Z., and Marinkovic, A. D. 2016. Surfacec Functionalization of Oxidized Multi-walled Carbon Nanotubes: Candida rugose Lipase Immobilization. Comptes Rendus Chimie. 19(3): 363-370. http://dx.doi.org/10.1016/j.crci.2015.10.008.
Verma, M. L., Naebe, M., Barrow, C. J., and Puri, M. 2013. Enzyme Immobilization on Amino-functionalized Multi-Walled Carbon Nanotubes: Structural and Biocatalytic Characterization. PLOS ONE. 8: 1-9.
Won, K., Kim, S., Kim, K.-J., Park, H. W., and Moon, S.-J. 2005. Optimization of Lipase Entrapment in ca-alginate Gel Beads. Process Biochemistry. 40(6): 2149-2154.
https://doi.org/10.1016/j.procbio.2004.08.014.
Gomez, J. M., Romero, M. D., and Fernandez, T. M. 2005. Immobilization of β-glucosidase on Carbon Nanotubes. Catalysis Letters. 101(3-4): 275-278.
Roy, S. K., Raha, S. K., Dey, S. K., and Chakrabarty, S. L. 1989. Immobilization of β-glucosidase from Myceliophthora thermophile D-14. Enzyme Enzyme and Microbial Technology. 11: 431-435.
Betigeri, S. S., and Neau, S. H. 2002. Immobilization of Lipase using Hydrophilic Polymers in the Form of Hydrogel Beads. Biomaterials. 23: 3627-3636.
https://doi.org/10.1016/S0142-9612(02)00095-9.
Knezevic, Z., Bobic, S., Milutinovic, A., Obradovic, B., Mojovic, L., and Bugraski, B. 2002. Alginate-immobilized Lipase by Electrostatic Extrusion for the Purpose of Palm Oil Hydrolysis in Lecithin/isooctane System. Process Biochemistry. 38: 313-318.
Keerti, A. K., Kumar, V., Dubey, A., and Verma, A. K. 2014. Kinetic Characterization and Effect of Immobilized Thermostable β-glucosidase in Alginate Gel Beads on Sugarcane Juice. ISRN Biochemistry. 2014: 178498. http://dx.doi.org/10.1155/2014/178498.
Mfombep, P. M., Seno, Z. N., and Isikhuemhen, O. S. 2013. Enzymatic Activities and Kinetic Properties of β-glucosidase from Selected White Rot Fungi. Advances in Biological Chemistry. 3:198-207.
Prlainovic, N. Z., Bezbradica, D. I., Knezevic-Jugovic, Z. D., Stevanovic, S. I., Avramov Ivic, M. L., Uskokovic, P. S., and Mijin, D. Z. 2013. Adsorption of Lipase from Candida Rugose on Multi Walled Carbon Nanotubes. Journal of Industrial and Engineering Chemistry. 19: 279-285.
Zniszczol, A., Herman, A. P., Szymanska, K., Mrowiec-Bialon, J., Walczak, K. Z., Jarzebski, A., and Boncel, S. 2016. Covalently Immobilized Lipase on Aminoalkyl-, Carboxy- and Hydroxyl-multi-wall Carbon Nanotubes in the Enantioselective Synthesis of Solketal Esters. Enzyme and Microbial Technology. 87-88: 61-69.
https://doi.org/10.1016/j.enzmictec.2016.02.015.
Dizge, N., Keskinler, B., and Tanriseven, A. 2008. Covalent Attachment of Microbial Lipase onto Microporous Styrene-divinylbenzene Copolymer by Means of Polyglutraldehyde. Colloids and Surfaces B: Biointerfaces. 66: 34-38.
Ortega, N, Busto, M. D., and Perez-Mateos, M. 1998. Optimization of β-glucosidase Entrapment in Alginate and Polyacrylamide Gels. Bioresource Technology. 64: 105-111.
Su, E., Xiaa, T., Gao, L., Dai, Q., and Zhang, Z. 2010. Immobilization of β-glucosidase and its Aroma-increasing Effect on Tea Beverage. Food and Bioproducts Processing. 88: 83-89.
Coutinho, T. C., Rojas, M. J., Tardioli, P. W., Paris, E. C., and Farinas, C. S. 2018. Nanoimmobilization of β-glucosidase onto Hydroxyapatite. International Journal of Biological Macromolecules. 119: 1042-1051 doi:10.1016/j.ijbiomac.2018.08.042.
Han, J., Luo, P., Wang, Y., Wang, L., Li, C., Zhang, W., Dong, J., and Ni, L. 2018. The Development of Nanobiocatalysis via the Immobilization of Cellulase on Composite Magnetic Nanomaterial for Enhanced Loading Capacity and Catalytic Activity. International Journal of Biological Macromolecules. 119: 692-700 doi:10.1016/j.ijbiomac.2018.07.176.
Maalim, K. Y. 2016. Optimization of β-glucosidase Enzyme Immobilisation on Ca-alginate and Carbon Nanotubes and the Evaluation of their Effectiveness Factors. Final Year Bachelor’s Degree Project Report. International Islamic University Malaysia, Kuala Lumpur.
Doran, P. M. 2013. Bioprocess Engineering Principles. 2nd Edition. Oxford: Academic Press.
Downloads
Published
Issue
Section
License
Copyright of articles that appear in Jurnal Teknologi belongs exclusively to Penerbit Universiti Teknologi Malaysia (Penerbit UTM Press). This copyright covers the rights to reproduce the article, including reprints, electronic reproductions, or any other reproductions of similar nature.
