Fig. 4

Formation of 11β-hydroxy-manoyl oxide by CYP76AH24. a Expression of CYP76AH24 in manoyl oxide-producing yeast cells resulted in the production of a new compound, which after isolation and structural analysis was identified as 11β-hydroxy-manoyl oxide. b GC–MS chromatogram of the products of an in vitro reaction using microsomal CYP76AH24 protein, manoyl oxide as substrate and NADPH as co-factor. A preparation of yeast microsomal membranes from a strain that expressed only CPR2 is used as control. Production of 11β-hydroxy-manoyl oxide was confirmed by comparison of retention time and mass spectrum with purified compound. c Mass spectrum of 11β-hydroxy-manoyl oxide standard (isolated from engineered yeast cells and characterized by NMR spectroscopy). d Mass spectrum of 11β-hydroxy-manoyl oxide produced by CYP76AH24 in an in vitro reaction