Figure 5

GC-MS analysis of Ss SS and Ss diTPS3 reaction products. Shown are total ion chromatograms (TIC) of reaction products from coupled enzyme assays with equal amounts of SsLPPS and either SsSS or SsdiTPS3 using GGPP 1 as a substrate. [A] Direct reaction products from coupled assays with SsLPPS and SsSS, [B] direct reaction products from coupled assays with SsLPPS and SsdiTPS3, [C] dephosphorylated reaction products of SsLPPS alone, [D] authentic manool standard (GlycoSyn, Gracefield, NZ), and [E] authentic sclareol standard (Sigma). GC-MS analysis was performed on an Agilent HP5ms column with electronic impact ionization at 70 eV. Results were confirmed with three independent experiments. 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.