In-depth understanding of a nano-bio interface between lysozyme and Au NP-immobilized N-doped reduced graphene oxide 2-D scaffolds.

In the present work, nitrogen-doped reduced graphene oxide (NrGO) was synthesized via a hydrothermal treatment of graphene oxide (GO) in the presence of urea. Gold nanoparticles (Au(0) NPs) were immobilized over the surface of NrGO (Au(0)-NrGO). Characterization of the Au(0)-NrGO nanocomposite via FT-IR spectroscopy, Raman spectroscopy, elemental mapping and XPS revealed the doping of N atoms during the reduction of GO. XRD and XPS studies confirmed the presence of Au(0) NPs and EDS analysis showed a 4.51 wt% loading of Au NPs in the Au(0)-NrGO nanocomposite. The morphology of Au(0)-NrGO was explored by SEM and TEM, which showed the presence of spherical Au metal NPs uniformly immobilized on the surface of NrGO. Further, studies on lysozyme (Lys) in the presence of Au(0)-NrGO by UV-visible, fluorescence, and circular dichroism spectroscopy revealed a conformational change in Lys and electrostatic interaction between Lys and Au(0)-NrGO. The DLS result showed an enhancement in the size of the Au(0)-NrGO and Lys conjugates. The Au(0)-NrGO-induced conformational changes in the structure of Lys resulted in a significant decrease in its activity at a certain concentration of Au(0)-NrGO. Moreover, the results showed that Lys favorably binds with the surface of Au(0)-NrGO, resulting in the formation of 2-D scaffolds possibly due to electrostatic and hydrophobic interactions, H-bonding, and interactions between the AuNPs and sulfur-containing amino acid residues of Lys. SEM exhibited the formation of conjugates in the form of 2-D scaffolds due to the biomolecular interactions between Lys and Au(0)-NrGO. The TEM studies revealed that Lys agglomerated around the Au(0) NPs immobilized on the surface of NrGO, which suggests the formation of a protein corona (PC) around the AuNPs. Furthermore, the favorable Au(0) NP-sulphur (PC) interaction was confirmed by the disappearance of the S-S stretching band in the Raman spectra. Overall, the results obtained provide insight into the nano-bio interface and formation of Au(0) NP-PC, which can be used for bioinspired applications, such as biosensing and imaging and the development of advanced functional Au NPs.