Abstract
Chitosanase could cleave β-1,4-linkage of chitosan to produce chitooligosaccharides (COS) with diverse biological activities. However, there are many limitations on the use of free chitosanase in industrial production. Enzyme immobilization is generally considered a valuable strategy in industrial-scale applications. In this study, the chitosanase Csn-BAC from Bacillus sp. MD-5 was immobilized on Fe3O4-SiO2 magnetic nanoparticles (MNPs) to enhance its properties, which could be recovered easily from reaction media using magnetic separation. The activities of Csn-BAC immobilized with MNPs (MNPs@Csn-BAC) were determined with temperature and pH, and the thermal- and pH-stabilities, respectively. The reusability of the MNPs@Csn-BAC was determined in repeated reaction cycles. Immobilization enhanced the thermal and pH stability of Csn-BAC compared with the free enzyme. After eight reaction cycles using MNPs@Csn-BAC, the residual enzyme activity was 72.15%. Finally, the amount of COS released by MNPs@Csn-BAC was 1.86 times higher than that of the free Csn-BAC in the catalytic performance experiment. The immobilized Csn-BAC exhibits broad application prospects in the production of COS.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Aggarwal, S., Chakravarty, A., and Ikram, S., 2021. A comprehensive review on incredible renewable carriers as promising platforms for enzyme immobilization & thereof strategies. International Journal of Biological Macromolecules, 167: 962–986, https://doi.org/10.1016/j.ijbiomac.2020.11.052.
Barbosa, O., Torres, R., Ortiz, C., Berenguer-Murcia, Á., Rodrigues R. C., and Fernandez-Lafuente R., 2013. Heterofunctional supports in enzyme immobilization: From traditional immobilization protocols to opportunities in tuning enzyme properties. Biomacromolecules, 14(8): 2433–2462, https://doi.org/10.1021/bm400762h.
Bayat, Z., Hassanshahian, M., and Cappello, S., 2015. Immobilization of microbes for bioremediation of crude oil polluted environments: A mini review. Open Microbiology Journal, 9: 48–54, https://doi.org/10.2174/1874285801509010048.
Bouabidi, Z. B., El-Naas, M. H., and Zhang, Z., 2018. Immobilization of microbial cells for the biotreatment of wastewater: A review. Environmental Chemistry Letters, 17: 241–257, https://doi.org/10.1007/s10311-018-0795-7.
Cao, S., Li, X., Lou, W., and Zong, M., 2014. Preparation of a novel magnetic cellulose nanocrystal and its efficient use for enzyme immobilization. Journal of Materials Chemistry B, 2: 5522–5530, https://doi.org/10.1039/c4tb00584h.
Cao, X., Xu, H., Li, F., Zou, Y., Ran, Y., Ma, X., et al., 2021. One-step direct transesterification of wet yeast for biodiesel production catalyzed by magnetic nanoparticle-immobilized lipase. Renewable Energy, 171: 11–21, https://doi.org/10.1016/j.renene.2021.02.065.
Du, G. H., Liu, Z. L., Xia, X., Chu, Q., and Zhang, S. M., 2006. Characterization and application of Fe3O4/SiO2 nanocomposites. Journal of Sol-Gel Science and Technology, 39(3): 285–291, https://doi.org/10.1007/s10971-006-7780-5.
Fang, G., Chen, H., Zhang, Y., and Chen, A., 2016. Immobilization of pectinase onto Fe3O4@SiO2-NH2 and its activity and stability. International Journal of Biological Macromolecules, 88: 189–195, https://doi.org/10.1016/j.ijbiomac.2016.03.059.
Hartmann, M., and Kostrov, X., 2013. Immobilization of enzymes on porous silicas-benefits and challenges. Chemical Society Reviews, 42(15): 6277–6289, https://doi.org/10.1039/c3cs60021a.
Jang, J. H., and Lim, H. B., 2010. Characterization and analytical application of surface modified magnetic nanoparticles. Microchemical Journal, 94(2): 148–158, https://doi.org/10.1016/j.microc.2009.10.011.
Khankari, S., Badoei-dalfard, A., and Karami, Z., 2021. Cross-linked enzyme aggregates of fibrinolytic protease BC1 immobilized on magnetic chitosan nanoparticles (CLEAs-FibmChi): Synthesis, purification, and characterization. Applied Biochemistry and Biotechnology, 193: 2004–2027, https://doi.org/10.1007/s12010-021-03494-z.
Li, Q., Chen, Y., Bai, S., Shao, X., Jiang, L., and Li, Q., 2020. Immobilized lipase in bio-based metal-organic frameworks constructed by biomimetic mineralization: A sustainable bio-catalyst for biodiesel synthesis. Colloids Surf B Biointerfaces, 188: 110812, https://doi.org/10.1016/j.colsurfb.2020.110812.
Muanprasat, C., and Chatsudthipong, V., 2017. Chitosan oligosaccharide: Biological activities and potential therapeutic applications. Pharmacology & Therapeutics, 170: 80–97, https://doi.org/10.1016/j.pharmthera.2016.10.013.
Nguyen, V. D., Styevkó, G., Madaras, E., Haktanirlar, G., Tran, A. T. M., Bujna, E., et al., 2019. Immobilization of β-galactosidase on chitosan-coated magnetic nanoparticles and its application for synthesis of lactulose-based galactooligosaccharides. Process Biochemistry, 84: 30–38, https://doi.org/10.1016/j.procbio.2019.05.021.
Pan, C. L., Hu, B., Li, W., Sun, Y., Ye, H., and Zeng, X. X., 2009. Novel and efficient method for immobilization and stabilization of β-D-galactosidase by covalent attachment onto magnetic Fe3O4-chitosan nanoparticles. Journal of Molecular Catalysis B Enzymatic, 61(3–4): 208–215, https://doi.org/10.1016/j.molcatb.2009.07.003.
Roger, S., and Sander, V. P., 2013. Enzyme immobilisation in biocatalysis: Why, what and how. Chemical Society Reviews, 42(15): 6223–6235, https://doi.org/10.1039/C3CS60075K.
Stöber, W., Fink, A., and Bohn, E., 1968. Controlled growth of monodisperse silica spheres in the micron size range. Journal of Colloid and Interface Science, 26(1): 62–69, https://doi.org/10.1016/0021-9797(68)90272-5.
Sun, H. H., Gao, L., Xue, C. H., and Mao, X. Z., 2020. Marine-polysaccharide degrading enzymes: Status and prospects. Comprehensive Reviews in Food Science and Food Safety, 19: 2767–2796, https://doi.org/10.1111/1541-4337.12630.
Thadathil, N., and Velappan, S. P., 2014. Recent developments in chitosanase research and its biotechnological applications: A review. Food Chemistry, 150: 392–399, https://doi.org/10.1016/j.foodchem.2013.10.083.
Unsoy, G., Yalcin, S., Khodadust, R., Gunduz, G., and Gunduz, U., 2012. Synthesis optimization and characterization of chitosan-coated iron oxide nanoparticles produced for biomedical applications. Journal of Nanoparticle Research, 14(11): 1–13, https://doi.org/10.1007/s11051-012-0964-8.
Wang, Q. D., Sun, J. A., Liu, Z., Huang, W. C., Xue, C. H., and Mao, X. Z., 2018. Coimmobilization of β-agarase and α-neoagarobiose hydrolase for enhancing the production of 3,6-anhydro-L-galactose. Journal of Agricultural and Food Chemistry, 66: 7087–7095, https://doi.org/10.1021/acs.jafc.8b01974.
Wang, W., Guo, N., Huang, W. C., Zhang, Z. H., and Mao, X. Z., 2018. Immobilization of chitosanases onto magnetic nanoparticles to enhance enzyme performance. Catalysts, 8(9): 401, https://doi.org/10.3390/catal8090401.
Xia, G., Cao, S., Xu, P., Li, X., Zhou, J., Zong, M., et al., 2017. Preparation of a nanobiocatalyst by efficiently immobilizing Aspergillus niger lipase onto magnetic metal-biomolecule frameworks (BioMOF). Chemcatchem, 9: 1794–1800, https://doi.org/10.1002/cctc.201700070.
Xie, W., and Zang, X., 2017. Covalent immobilization of lipase onto aminopropyl-functionalized hydroxyapatite-encap-sulated-γ-Fe2O3 nanoparticles: A magnetic biocatalyst for inter-esterification of soybean oil. Food Chemistry, 227: 397–403, https://doi.org/10.1016/j.foodchem.2017.01.082.
Xie, W., and Zang, X., 2018. Lipase immobilized on ionic liquid-functionalized magnetic silica composites as a magnetic biocatalyst for production of trans-free plastic fats. Food Chemistry, 257: 15–22, https://doi.org/10.1016/j.foodchem.2018.03.010.
Yang, G. S., Sun, H. H., Cao, R., Liu, Q., and Mao, X. Z., 2020. Characterization of a novel glycoside hydrolase family 46 chitosanase, Csn-BAC, from Bacillus sp. MD-5. International Journal of Biological Macromolecules, 146: 518–523, https://doi.org/10.1016/j.ijbiomac.2020.01.031.
Zhang, L. Y., Zhu, X. J., Sun, H. W., Chi, G. R., Xu, J. X., and Sun, Y. L., 2010. Control synthesis of magnetic Fe3O4-chitosan nanoparticles under UV irradiation in aqueous system. Current Applied Physics, 10(3): 828–833, https://doi.org/10.1016/j.cap.2009.10.002.
Acknowledgements
This work was supported by the National Key Research and Development Program of China (No. 2019 YFD0901902), the National Natural Science Foundation of China (No. 31801574), the Central Public-interest Scientific Institution Basal Research Fund, YSFRI, CAFS (No. 20603022021020), and Qingdao Science and Technology Demonstration and Guidance Project for Benefiting the People (No. 20-3-4-28-nsh).
Author information
Authors and Affiliations
Corresponding authors
Rights and permissions
About this article
Cite this article
Cheng, Y., Li, Z., Sun, H. et al. Immobilization of Chitosanase on Magnetic Nanoparticles: Preparation, Characterization and Properties. J. Ocean Univ. China 21, 1381–1388 (2022). https://doi.org/10.1007/s11802-022-5189-6
Received:
Revised:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s11802-022-5189-6