The β-1,3-glucanase gene in Ostrinia furnacalis was first obtained by RT-PCR. The real-time fluorescence quantitative PCR showed that the expression level of β-1,3-glucanase in the midgut of O. furnacalis was higher than in other tissues. Moreover, the expression level in the larval stage was higher in egg, pupa, and adult stages. The optimal pH of recombinant O. furnacalis β-1,3-glucanase OfLam to the substrate laminarin was 4.5, and the optimum reaction temperature was 50°C. The enzyme exhibited a KM of 1.59 ± 0.28 mg/mL and a kcat of 15.8 ± 0.66 s-1 . Ostrinia furnacalis β-1,3-glucanase has a similar catalytic efficiency to other insect-derived β-1,3-glucanases. The recombinant OfLam has a broad substrate spectrum and can hydrolyze fungal cell walls, suggesting a new source of enzymes for biological control strategies that target fungal cell walls.