The preparation of functional materials and value-added chemicals using biomass-derived feedstocks is an interesting topic. Herein, we propose a method to prepare Ru/graphene catalyst using glucose as the carbon source. The catalyst was characterized by scanning and transmission electron microscopies, Raman and X-ray photoelectron spectroscopies, and X-ray diffraction. It was found that Ru particles of 2-6 nm were supported uniformly on high-quality graphene. The performance of the Ru/graphene for hydrogenation of levulinic acid to γ-valerolactone was studied. The Ru/graphene had much higher activity for the hydrogenation reaction than commercial Ru/C, and was active even at room temperature. The high activity was partially due to the unique structure of graphene, which resulted in an electron-enriched Ru nanocatalyst. In addition, the Ru/graphene could be reused at least four times without considerable decrease in activity, indicating the excellent stability of the catalyst under our conditions.