Covalently functionalized single-walled carbon nanotubes at reverse micellar interface: a strategy to improve lipase activity

Abstract

The present work reports covalent functionalization of single-walled carbon nanotubes (f-SWNTs) to introduce hydrophilicity to the otherwise amphiphobic nanotubes. The charge and spacer length of the functional moiety were varied by using quaternized ethylene diamine, 6-aminocaproate, quaternized (ethylenedioxy)bis(ethylamine), and a poly(ethylene glycol) (PEG) unit (f-SWNT-1 to f-SWNT-4, respectively). These f-SWNTs with varying degrees of hydrophilicity were incorporated within cetyltrimethyl ammonium bromide (CTAB) reverse micelles to develop stable self-assembled nanohybrids. An optimum hydrophilicity on the SWNT surface led to interfacial localization of f-SWNTs resulting in the augmentation of space at the interface. A surface-active enzyme, lipase, localized at this enhanced interface of f-SWNT-containing CTAB reverse micelles exhibited significant activation (2.5-fold) compared to that in the absence of the nanoconstructs. This improvement in lipase activity was mainly due to the smooth occupancy of lipase and also presumably because of the increase in the concentrations of both substrate and the enzyme at the augmented interface. Interestingly, the f-SWNTs that activate lipase in reverse micelles deactivate the same enzyme in water. The dispersion of f-SWNTs in water and its matching integration at the interface of reverse micelles were confirmed through transmission electron microscopic (TEM) investigations. The interfacial localization of these nanoconstructs was also established from the distinct fluorescence behavior of a hydrophobic fluorescent probe, fluorescein isothiocyanate (FITC), adsorbed onto the f-SWNT surface. In concurrence with the observed lipase activity, the corresponding changes in the enzyme conformation within f-SWNTs integrated reverse micelle as well as in aqueous medium were studied by circular dichroism (CD) and Fourier transform infrared (FTIR) spectroscopy.