BackgroundThis study provides fundamental information on the influence of graphene oxide (GO) nanosheets and glycans on protein catalytic activity, dynamics, and thermal stability. We provide evidence of protein stabilization by glycans and how this strategy could be implemented when GO nanosheets is used as protein immobilization matrix. A series of bioconjugates was constructed using two different strategies: adsorbing or covalently attaching native and glycosylated bilirubin oxidase (BOD) to GO.ResultsBioconjugate formation was followed by FT-IR, zeta-potential, and X-ray photoelectron spectroscopy measurements. Enzyme kinetic parameters (km and kcat) revealed that the substrate binding affinity was not affected by glycosylation and immobilization on GO, but the rate of enzyme catalysis was reduced. Structural analysis by circular dichroism showed that glycosylation did not affect the tertiary or the secondary structure of BOD. However, GO produced slight changes in the secondary structure. To shed light into the biophysical consequence of protein glycosylation and protein immobilization on GO nanosheets, we studied structural protein dynamical changes by FT-IR H/D exchange and thermal inactivation.ConclusionsIt was found that glycosylation caused a reduction in structural dynamics that resulted in an increase in thermostability and a decrease in the catalytic activity for both, glycoconjugate and immobilized enzyme. These results establish the usefulness of chemical glycosylation to modulate protein structural dynamics and stability to develop a more stable GO-protein matrix.Electronic supplementary materialThe online version of this article (doi:10.1186/s12951-015-0134-0) contains supplementary material, which is available to authorized users.

We constructed a biosensor by electrodeposition of gold nano-particles (AuNPs) on glassy carbon (GC) and subsequent formation of a 4-mercaptobenzoic acid self-assembled monolayer (SAM). The enzyme horseradish peroxidase (HRP) was then covalently immobilized onto the SAM. Two forms of HRP were employed: non-modified and chemically glycosylated with lactose. Circular dichroism (CD) spectra showed that chemical glycosylation did neither change the tertiary structure of HRP nor the heme environment. The highest sensitivity of the biosensor to hydroquinone was obtained for the biosensor with HRP-lactose 1 (414 nA μM−1 ) compared to 378 nA μM−1 for the one employing non-modified HRP. The chemically glycosylated form of the enzyme catalyzed the reduction of hydroquinone more rapidly than the native form of the enzyme. The sensor employing lactose-modified HRP also had a lower limit of detection (74 μM) than the HRP biosensor (83 μM). However, most importantly, chemically glycosylation improved the long-term stability of the biosensor, which retained 60% of its activity over a four-month storage period compared to only 10% for HRP. These results highlight improvements by an innovative stabilization method when compared to previously reported enzyme-based biosensors.

Graphene possesses excellent mechanical strength and chemical inertness with high intrinsic carrier mobility and superior flexibility making them exceptional candidates for optoelectronic applications. Graphene quantum dots (GQDs) derived from graphene domains have been widely explored to study their photoluminescence properties which can be tuned by size. GQDs are biocompatible, low cytotoxic, strongly luminescent and disperse well in polar and non-polar solvents showing bright promise for the integration into devices for bioimaging, light emitting and photovoltaic applications. In the present study, graphene quantum dots were synthesized by an electrochemical cyclic voltammetry technique using reduced graphene oxide (rGO). GQDs have been incorporated into binder free TiO2 paste and studied as a photoelectrode material fabricated on ITO/PEN substrates for flexible dye sensitized solar cells (DSSCs). DSSC based on GQDs-TiO2 exhibited open circuit output potential difference (Voc) of 0.73 V, and short circuit current density (Jsc) of 11.54 mA cm-2 with an increment in power conversion efficiency by 5.48 %, when compared with those with DSSC build with just a TiO2 photoanode (open-circuit output potential difference (Voc) of 0.68 V and short circuit density (Jsc) of 10.67 mA cm-2). The results have been understood in terms of increased charge extraction and reduced recombination losses upon GQDs incorporation.

No abstract

No abstract