A globular protein, β-lactoglobulin (BLG), was used to decorate reduced graphene oxide sheets (RGO) and the obtained BLG-RGO composite can be dispersed in aqueous solution with pH-sensitive solubility. The morphology of the BLG-RGO composite was studied by transmission electron microscopy (TEM) and atomic force microscopy (AFM). The results indicate that BLG-RGO is effectively exfoliated with an average thickness of 2.5 nm. UV-vis spectra were performed to examine the reduction degree and determine the optimum concentration of β-lactoglobulin and appropriate pH value. Furthermore, Raman spectra demonstrate that β-Lactoglobulin promotes the chemical reduction process of graphene oxide and benefits to repair the crystal defects. Due to the adsorption of β-Lactoglobulin on the surface of graphene sheets, the BLG-RGO composite was further used as template for Au nanoparticles assembly. These Au nanoparticles assembled on the BLG-RGO composite were shown to yield a large SERS enhancement for Rhodamine 6G.