Unprecedented Cyclization Catalyzed by a Cytochrome P450 in Benzastatin Biosynthesis TsutsumiHayama KatsuyamaYohei IzumikawaMiho TakagiMotoki FujieManabu SatohNoriyuki Shin-yaKazuo OhnishiYasuo 2018 Benzastatins have unique structures probably derived from geranylated <i>p</i>-aminobenzoic acids. The indoline and tetrahydroquinoline scaffolds are presumably formed by cyclization of the geranyl moiety, but the cyclization mechanism was unknown. We studied the benzastatin biosynthetic gene cluster of <i>Streptomyces</i> sp. RI18; functions of the six enzymes encoded by it were analyzed by gene disruption in a heterologous host and in vitro enzyme assays. We propose the biosynthetic pathway for benzastatins in which a cytochrome P450 (BezE) is responsible for the cyclization of geranylated <i>p</i>-acetoxyaminobenzoic acids; BezE catalyzes elimination of acetic acid to form an iron nitrenoid, nitrene transfer to form an aziridine ring, and nucleophilic addition of hydroxide ion to C-10 and chloride ion to C-9 to generate the indoline and tetrahydroquinoline scaffolds, respectively. Discovery of this enzyme, which should be termed cytochrome P450 nitrene transferase, provides an important insight into the functional diversity of cytochrome P450.