Two different kinds of ZnO nanoparticles (NPs) with distinct particle sizes were synthesized by a precipitation method. For the first time, the effects of two distinct sizes ZnO NPs on the growth, photoluminescence properties and kojic acid (KA) yield of Aspergillus oryzae (A. oryzae) were examined. The findings showed that ZnO-A and -B NPs, with average particle sizes of 11.93 nm and 34.41[Formula: see text]nm, respectively, exhibited a hexagonal crystal structure. It was found that when the concentration of NPs was below 100[Formula: see text][Formula: see text]g/mL, ZnO NPs were nontoxic and served as activators to stimulate the growth of A. oryzae, and the biomass of solid and liquid medium, the total integral emission intensity, the content of extracellular protein and total organic carbon and KA yield of A. oryzae grown with ZnO-A (ZnO-B) NPs were about 2.19 (1.97), 2.47 (1.56), 1.67 (1.16), 2.45 (1.95), 0.47 (0.50) and 2.07 (1.37) times more than that of control sample (without NPs), respectively.