Figure 6

Production of sclareol in engineered yeast cells. Shown are GC-MS total ion chromatograms (TIC) of extracts from engineered yeast strains expressing [A] yeast GGPP synthase (ScGGPPS), [B] ScGGPPS and SsLPPS, [C-D] ScGGPPS, SsLPPS and SsSS, and [E-F] ScGGPPS, SsLPPS and SsSdiTPS3.Sclareol was detected at similar levels in both the media [D &F] and harvested yeast cells [C &E] of each yeast culture, with a lower amount of impurities being observed in the media. GC-MS analysis was performed on an Agilent HP5ms column with electronic impact ionization at 70 eV. Identification of reaction products was achieved by comparison to authentic standards or reference mass spectra from the National Institute of Standards and Technology MS library searches (Wiley W9N08L): peak a, epi-manoyl oxide 8; peak b, manoyl oxide 7; peak c, putative 13(16),-14-labdien-8-ol; peak d, putative copalol; peak e, sclareol 4; peak f, unknown compound; peak g, labda-13-en-8,15-diol; peak i, manool 6.