Manoyl Oxide (13R), the Biosynthetic Precursor of Forskolin, Is Synthesized in Specialized Root Cork Cells in Coleus forskohlii

Expanding the Landscape of Diterpene Structural Diversity through Stereochemically Controlled Combinatorial Biosynthesis

“…Generation of expression constructs, expression in N. benthamiana , S. cerevisiae and E. coli , extraction analysis of products, and structural elucidation by NMR were done as previously described and are given in supplementary material and methods 6a, 19…”

“…Cf= C. forskohlii ; Ss= S. sclarea ; Mv= M. vulgare ; Ep= Euphorbia peplus ; Os= O. sativa ; Tw= T. wilfordii ; Zm= Z. mays . Diterpenes in samples expressing solely class II diTPSs are likely the result of endogenous phosphatase activity and non‐specific rearrangement (Figure S7) 6a, 7…”

“…(13 R )‐(+)‐manoyl oxide ( 16 a ), a precursor for the diterpenoid pharmaceutical forskolin,6a was formed from (+)‐8‐LPP by the combination of Salvia sclarea SsLPPS with Marrubium vulgare MvTPS5 at 3.7 times higher levels than the native combination of CfTPS2 and CfTPS3 from Coleus forskohlii , which may be relevant for future production. Stereoselective access to manoyl oxide stereoisomers is a prerequisite for the biotechnological production of forskolin derivatives with novel stereochemical configurations.…”

“…Several of the combinations provided skeletons of high‐value diterpenoids, for example, miltiradiene ( 25 ), the backbone of cardioprotectives (tanshinones)15 and antioxidants (carnosoic acid),16 16 a and 23 a as precursors for the drug forskolin,6a and the synthetic ambergris substitute (−)‐ambrox, which is used in high‐end fragrances 17. For stereoselective production of these targets, expression‐optimized variants of CfTPS1/CfTPS3, CfTPS2/CfTPS3, and TwTPS21/EpTPS1 were introduced stably into the genome of a S. cerevisiae strain engineered for diterpene production.…”

Functional divergence of diterpene syntheses in the medicinal plant Salvia miltiorrhiza Bunge

“…Miltiradiene is a precursor to at least cryptotanshinone (Guo et al, 2013), and RNA interference (RNAi) knockdown of SmCPS1 expression reduces tanshinone production, at least in hairy root cultures (Cheng et al, 2014). The identification of SmCPS1 and SmKSL1 has been followed by that of many related diTPSs from other Lamiaceae plant species (Caniard et al, 2012;Sallaud et al, 2012;Schalk et al, 2012;Brückner et al, 2014;Pateraki et al, 2014). These largely exhibit analogous activity, particularly the CPSs, which produce CPP or the stereochemically related 8a-hydroxy-labd-13E-en-15-yl diphosphate (LDPP) rather than the enantiomeric (ent) CPP relevant to GA biosynthesis.…”