Unprecedentedly, an eco-friendly green approach is used to reduce graphene oxide via two bacterial strains (Streptococcus thermophilus and Bacillus coagulans). Bionanotechnology and microbial reduction of GO are considered safe for the environment, cheap, easy, and green. The UV-visible spectroscopy of rGO nanoparticles revealed a prominent absorption peak at 260 nm attributed to redshift. SEM scan revealed a wavy fold and semispherical nanoparticles distributed unevenly with sharp edges and dimensions ranging between (22-28nm). The peaks of oxygen functional groups were reduced in the FTIR spectra; thus a minor peak was detected at wavelength 3050 indicates there is a minor degradation or aggregation of rGO. X-ray diffraction measurements exhibited sharp signals, which indicates a crystal shape of rGO and the diffraction peak appears at 2θ = 28.21° corresponded to a d-spacing roughly 0.31nm. Finally, AFM images exhibited the roughness of rGO nanoparticles. Current results revealed the fruitful bacterial reduction of GO by the exclusion of water molecules and efficient oxygen groups from the graphene oxide interlayer.