Mechanistic Characterisation and Engineering of Sesterviolene Synthase from Streptomyces violens

The sesterviolene synthase from Streptomyces violens was identified and represents the second known sesterterpene synthase from bacteria. Isotopic labelling experiments in conjunction with DFT calculations were performed that provided detailed insight into its complex cyclisation mechanism. Enzyme engineering through site‐directed mutagenesis gave access to a high‐yielding enzyme variant that provided six additional minor products and the main product in sufficient quantities to study its chemistry.