The present disclosure provides an in vitro method for producing a cannabinoid, the method comprising reacting a N prenylated aromatic compound with a flavin-dependent oxidase in a reaction mixture to form the cannabinoid, wherein the flavin-dependent oxidase is not derived from C. sativa.

The application relates to recombinant polypeptides having berberine bridge enzyme (BBE) activity that are derived from microorganisms, such as Phytohabitans sufuscus, Streptomyces sp. AJS327, Streptomyces varsoviensis, Actinomadura pelletieri, Streptomyces flaveolus, Streptomyces sp. Ru71. or Streptomyces sp. CNH287, and the use of these recombinant polypeptides in compositions and methods for the oxidative cyclization of prenylated compounds, such as CBGA, in carrying out the biosynthesis, including cell-free biosynthesis, of cyclized cannabinoid compounds, such as CBCA, THCA, and CBDA.