Novel reduced graphene oxide-supported palladium nanoparticles (RGO-PN) were synthesized under ultrasonication, a method that utilizes Coleus amboinicus as a bio-reduction agent. Green synthesized RGO-PN nanoparticles with a crystallite size in the range of 40–50 nm were confirmed in X-ray diffraction (XRD) spectra. RGO-PN show an absorption peak at 220 nm while reduced graphene oxide (RGO) shows its maximal absorbance at 210 nm. The scanning electron microscope image revealed that 40-nm-sized spherical-shaped palladium nanoparticles stick well to reduced graphene oxide sheets, which is consistent and correlated well with the XRD pattern. Moreover, a high-resolution morphological image of RGO-PN100 was obtained by TEM analysis, which shows the anchoring of palladium nanoparticles (PN) on RGO nanosheets. Green synthesized RGO-PN100 nanoparticles from Coleus amboinicus show better reduction kinetics for 4-nitrophenol at 40 min, suggesting that RGO-PN prepared from Coleus amboinicus serve as an excellent catalytic reducing agent. Furthermore, they show remarkable antibacterial activity against Escherichia coli (ATCC 25922). Thus, green synthesized RGO-supported palladium nanoparticles demonstrated that enhanced catalytic activity and antibacterial activity both play an important role in the environmental and medical disciplines.