Spinosad, extracted from Saccharopolyspora spinosa, one of the most successful commercial bio-insecticides, is derived from a family of macrocyclic lactones. It shows excellent potent insecticidal activities, low residue, and low environmental effect. Here, we reviewed the biosynthetic pathway of spinosad, the chemoenzymatic method of spinosad synthesis, and the strain improvement method and the heterologous expression of spinosad.