Abstract
Silk fibroin derived from Bombyx mori is a biomacromolecular protein with excellent biocompatibility. The aim of this work was to develop silk fibroin nanoparticles (SFNs) derived from the fibrous protein, which is a novel vector for enzyme modification in food processing. Silk fibroin was dissolved in highly concentrated CaCl2 and subjected to lengthy desalting in water. The resulting liquid silk, which contained water-soluble polypeptides with molecular mass ranging from 10 to 200 kDa, and β-glucosidase were added rapidly into acetone. The β-glucosidase molecules were embedded into silk fibroin nanoparticles, forming β-glucosidase–silk fibroin nanoparticles (βG–SFNs) with a diameter of 50–150 nm. The enzyme activity of the βG–SFN bioconjugates was determined with p-nitrophenyl-β-d-glucoside as the substrate, and the optimum conditions for the preparation of βG–SFNs were investigated. The enzyme activity recovery of βG–SFNs was 59.2 % compared to the free enzyme (specific activity was 1 U mg-1). The kinetic parameters of the βG–SFNs and the free β-glucosidase were the same. The βG–SFNs had good operational stability and could be used repeatedly. These results confirmed that silk protein nanoparticles were good carriers as bioconjugates for the modification of enzymes with potential value for research and development. The method used in this study has potential applications in food processing and the production of flavour agents.
Similar content being viewed by others
References
D. San. (1992). International Union of Biochemistry. Academic Press, 310
Krammer, G., Winterhalter, P., Schwab, M., et al. (1991). Journal of Agricultural and Food Chemistry, 39, 778–781.
Gunata, Z. Y., Bayonove, C. T., Tapiero, R. E., et al. (1990). Journal of Agricultural and Food Chemistry, 38, 1232–1236.
Hang, H. T (1995). Agriculture. Food Chemistry, 3, 141–146.
Martino, A., Pifferi, P. G., & Spagna, G. (1994). Journal of Chemical Technology and Biotechnology, 60, 247–252.
Nicolus, P., Racetz, E., Sauvageat, J. (1997). European Patent. EPO777972 AL1997-10-H0248.
Zhang, Y. Q. (1998). Biotechnology Advances, 16, 961–971.
Zhang,Y. Q., Zhu, J., Shen, W. D. (1997). Journal of Soochow University, 13, 118–125.
Zhang, Y. Q., Zhu, J., & Gu, R. A. (1998). Applied Biochemistry and Biotechnology, 75, 215–233.
Zhang, Y. Q., & Zhou, Y. Z. (1995). Progress in Biochemistry and Biophysics, 22(2), 162–165.
Zhang, Y. Q., Zhou, Y. Z. (1995). Journal of Suzhou Institute Silk Textile Technology, 15, 99–103.
Zhang, Y. Q., Gu, R. A., Zhu, J., & Cao, P. G. (1998). Progress in Biochemistry and Biophysics, 25(3), 275–278.
Zhang, Y. Q., Shen, W. D., Gu., R. A., et al. (1998). Analytica Chimica Acta, 369, 123–128.
Miyairi, S., Sugiura, M., Fukui, S. (1978). Agricultural and Biological Chemistry, 42(9), 1661–1667.
Zhang, Z. Z., Li, Y. B., Su, E. Z., Li, P.(2004). Food Fermentation Industry, 30(6), 6–9.
Zhang, Y. Q., Tao, M. L., Shen, W. D., et al. (2004). Biomaterials, 25, 3751–3759.
Zhang, Y. Q., Zhou, W. L., Shen, W. D., et al. (2005) Journal of Biotechnology, 120, 315–326.
Zhang, Y. Q., Ma, Y., Xia, Y. Y., et al. (2006). Journal of Controlled Release, 115, 307–315.
Zhang, Y. Q., Ma, Y., Xia,Y. Y., et al. (2006) Journal of Biomedical Materials Research Part B: Applied Biomaterials, 79B, 275–283.
Zhang, Y. Q., Shen, W. D., Xiang, R. L., et al. (2007). Journal of Nanoparticle Research, 9, 885–900.
Yan, H. B., Zhang, Y. Q., Ma, Y. L., & Zhou, L. X. (2009). Journal of Nanoparticle Research, 11, 1937–1946.
Zhang, Y. Q., Xiang, R. L., Yan, H. B., & Chen, X. X. (2008). Chemical Journal of Chinese Universities, 29, 628–633.
Zhou, Z. Z., Zhang, Y.Q. (2011). Advances in Materials Research, 175–176, 186–191.
Zhang, Y. Q. (2007). Silk fibroin nanoparticles fixed with enzyme and their production. PCT, PCT/CN2007/001032.
Su, E., Xia, T., Gao, L., Dai, Q., & Zhang, Z. (2010). Food and Bioproducts Processing, 88(2–3), 83–89.
Tu, M. B., Zhang, X., Kurabi, A., et al. (2006). Biotechnological Letters, 28, 151–156.
Serefoglou, E., Litina, K., Gournis, D., et al. (2008). Chemistry of Materials, 20, 4106–4115.
Acknowledgments
The authors gratefully acknowledge the earmarked fund (CARS-22-ZJ0504) for China Agriculture Research System (CARS) and a project funded by the Priority Academic Program Development of Jiangsu Higher Education Institutions, People’s Republic of China.
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Cao, TT., Zhou, ZZ. & Zhang, YQ. Processing of β-Glucosidase–Silk Fibroin Nanoparticle Bioconjugates and Their Characteristics. Appl Biochem Biotechnol 173, 544–551 (2014). https://doi.org/10.1007/s12010-014-0861-y
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s12010-014-0861-y