Abstract
Common enzyme immobilization methods on nanomaterials (adsorption, covalent binding, crosslinking, encapsulation) often generate problems in enzyme leaching, 3D structure change and diffusion resistance. We show here a detailed site-specific enzyme immobilization method that overcomes the foresaid limitations. It is based on the specific interaction between His-tagged enzyme and single-walled carbon nanotubes modified with N α ,N α-bis(carboxymethyl)-l-lysine hydrate. This method does not require enzyme purification and the resulting nanoscale biocatalyst can maintain high enzyme activity and stability. The enzyme-loading capacity is also comparable with the reported immobilization capacity on carbon nanotubes by either covalent binding or adsorption. Furthermore, the immobilization is reversible for several cycles while maintaining high enzyme activity and the nanoscale biocatalyst can be regenerated easily.
This is a preview of subscription content, log in via an institution to check access.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Wang, P. (2006) Nanoscale biocatalyst systems. Curr. Opin. Biotechnol. 17, 574–579.
Asuri, P., Karajanagi, S. S., Dordick, J. S., and Kane, R. S. (2006) Directed assembly of carbon nanotubes at liquid-liquid interfaces: Nanoscale conveyors for interfacial biocatalysis. J. Am. Chem. Soc. 128, 1046–1047.
Zhao, X., Jia, H., Kim, J., and Wang, P. (2009) Kinetic limitations of a bioelectrochemical electrode using carbon nanotube-attached glucose oxidase for biofuel cells. Biotechnol. Bioeng. 104, 1068–1074.
Kim, M. I., Kim, J., Lee, J., Jia, H., Bin Na, H., Youn, J. K., Kwak, J. H., Dohnalkova, A., Grate, J. W., Wang, P., Hyeon, T., Park, H. G., and Chang, H. N. (2007) Crosslinked enzyme aggregates in hierarchically-ordered mesoporous silica: A simple and effective method for enzyme stabilization. Biotechnol. Bioeng. 96, 210–218.
Elgren, T. E., Zadvorny, O. A., Brecht, E., Douglas, T., Zorin, N. A., Maroney, M. J., and Peters, J. W. (2005) Immobilization of active hydrogenases by encapsulation in polymeric porous gels. Nano Lett. 5, 2085–2087.
Zhu, Y., Kaskel, S., Shi, J., Wage, T., and van Pee, K. H. (2007) Immobilization of Trametes versicolor laccase on magnetically separable mesoporous silica spheres. Chem. Mat. 19, 6408–6413.
Asuri, P., Bale, S. S., Pangule, R. C., Shah, D. A., Kane, R. S., and Dordick, J. S. (2007) Structure, function, and stability of enzymes covalently attached to single-walled carbon nanotubes. Langmuir 23, 12318–12321.
Hudson, S., Cooney, J., and Magner, E. (2008) Proteins in mesoporous silicates. Angew. Chem. Int. Ed. 47, 8582–8594.
Asuri, P., Karajanagi, S. S., Yang, H. C., Yim, T. J., Kane, R. S., and Dordick, J. S. (2006) Increasing protein stability through control of the nanoscale environment. Langmuir 22, 5833–5836.
Yim, T. J., Liu, J. W., Lu, Y., Kane, R. S., and Dordick, J. S. (2005) Highly active and stable DNAzyme–carbon nanotube hybrids. J. Am. Chem. Soc. 127, 12200–12201.
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2011 Springer Science+Business Media, LLC
About this protocol
Cite this protocol
Wang, L., Jiang, R. (2011). Reversible His-Tagged Enzyme Immobilization on Functionalized Carbon Nanotubes as Nanoscale Biocatalyst. In: Wang, P. (eds) Nanoscale Biocatalysis. Methods in Molecular Biology, vol 743. Humana Press. https://doi.org/10.1007/978-1-61779-132-1_8
Download citation
DOI: https://doi.org/10.1007/978-1-61779-132-1_8
Published:
Publisher Name: Humana Press
Print ISBN: 978-1-61779-131-4
Online ISBN: 978-1-61779-132-1
eBook Packages: Springer Protocols