Abstract
Diterpenoid natural products cover a vast chemical diversity and include many medicinally and industrially relevant compounds. All diterpenoids derive from a common substrate, (E,E,E)-geranylgeranyl diphosphate, which is cyclized into one of many scaffolds by a diterpene synthase (DTS). While diterpene biosynthesis has been extensively studied in plants and fungi, bacteria are now recognized for their production of unique diterpenoids and are likely to harbor an underexplored reservoir of new DTSs. Bacterial diterpenoid biosynthesis can be exploited for the discovery of new natural products, a better mechanistic understanding of DTSs, and the rational engineering of whole metabolic pathways. This chapter describes methods and protocols for identification and characterization of bacterial DTSs, based on our recent work with the DTSs involved in platensimycin and platencin biosynthesis.