Isonitrile group has been identified in many natural products.Due to the broad reactivity of N=C triple bond,these natural products have valuable pharmaceutical potentials.This review summarizes the current biosynthetic pathways and the corresponding enzymes that are responsible for isonitrile-containing natural product generation.Based on the strategies utilized,two fundamentally distinctive approaches are discussed.In addition,recent progress in elucidating isonitrile group formation mechanisms is also presented.