Prenylated isoflavonoids have excellent biological activity and are widely used in pharmaceutical and food industry.Prenyl unit is one of the pharmacophores of prenylated isoflavonoids.Chemical prenylation usually has poor chemoselectivity and regioselectivity for several positions,and protection and deprotection of hydroxyl are usually necessary in the synthetic route.In plants,membrane-bound prenyltransferases transfer the prenyl unit from dimethylallyl diphosphate to the isoflavonoid scaffold.Prenyltransferases from plants are membrane-bound proteins and catalytic activities of these proteins are very low,and these proteins are difficult to overproduce and purify than soluble enzymes.Prenyltransferases from fungi are soluble and exhibit promiscuity toward diverse aromatic acceptors and prenyl donors,which have great potential to be biocatalysts,and these prenyltransferases accept isoflavonoids as subustrates.The present review deals with the structural characteristics of prenylated isoflavonoids and the prenylation of isoflavonoids by prenyltransferases of isoflavonoids in the past 20 years.