Modular Chemoenzymatic Synthesis of Terpenes and their Analogues

Abstract

Non-natural terpenoids offer potential as pharmaceuticals and agrochemicals. However, their chemical syntheses are often long, complex, and not easily amenable to large-scale production. Herein, we report a modular chemoenzymatic approach to synthesize terpene analogues from diphosphorylated precursors produced in quantitative yields. Through the addition of prenyl transferases, farnesyl diphosphates, (2E,6E)-FDP and (2Z,6Z)-FDP, were isolated in greater than 80 % yields. The synthesis of 14,15-dimethyl-FDP, 12methyl-FDP, 12-hydroxy-FDP, homo-FDP, and 15-methyl-FDP was also achieved. These modified diphosphates were used with terpene synthases to produce the unnatural sesquiterpenoid semiochemicals (S)-14,15-dimethylgermacrene D and (S)-12-methylgermacrene D as well as dihydroartemisinic aldehyde. This approach is applicable to the synthesis of many non-natural terpenoids, offering a scalable route free from repeated chain extensions and capricious chemical phosphorylation reactions.

Alternative metabolic pathways and strategies to high-titre terpenoid production inEscherichia coli

“…A novel and simpler three-step pathway which uses isoprenol or prenol as feedstocks to produce IPP and DMAPP was recently exploited by several groups. [129][130][131][132][133][134] This pathway decouples terpenoid biosynthesis from central metabolism and therefore has the potential to increase metabolic ux towards the production of terpenoids. 135 The pathway also reduces any negative impacts on cell growth, since glucose (or other carbon source used for bacterial growth) is reserved for primary metabolism.…”

The Terpene Mini-Path, a New Promising Alternative for Terpenoids Bio-Production

“…The presence of both ThiM and IP kinases in E. coli could, by analogy, explain the reported ability of some microorganisms to use added DMAOH and IOH as substrates for the generation of DMAPP/IPP, as shown above. ThiM kinase was also proved to be useful for IOH as a substrate and was, therefore, combined with IPKs to produce the universal diphosphates in vitro [ 29 , 30 ] or in vivo [ 13 ] in order to generate various terpenoids using DMAOH/IOH as precursors.…”

“…In 2020, two papers appeared, both taking advantage of the use of another true kinase (the ThiM kinase from E. coli , see above) to phosphorylate DMAOH and IOH into DMAPP and IPP in conjunction with the Methanocaldococcus jannaschii IPK [ 29 , 30 ]. In the former case, the formed DMAPP and IPP were used as substrates of different linear prenyl transferases ( Scheme 6 ) to access ( E , E )-FPP, ( Z , Z )-FPP, GPP, and GGPP.…”

“…The TMP has also been exploited to generate, from homologs of DMAOH and IOH, various linear diphosphates homologous to FPP. Some of them were then used as substrates for germacrene D and amorphadiene sesquiterpene synthases, providing the corresponding unnatural terpenoids [ 30 ].…”

Nature Chose Phosphates and Chemists Should Too: How Emerging P(V) Methods Can Augment Existing Strategies

“…The elementary enzymatic steps of terpene and steroid biosynthesis involve the assembly-line-like construction of polyunsaturated systems from a simple building block: dimethylallylpyrophosphate (DMAPP). 8 The preorganized generation of carbocation intermediates, driven by pyrophosphate formation, allows for the controlled creation of C—C bonds that is currently outside the realm of chemical synthesis in the laboratory. In contrast, organic chemists generally use classical leaving groups and multistep sequences involving protecting group chemistry to accomplish the same overall construction.…”