Within and beyond organelle engineering: strategies for increased terpene production in yeasts and plants

Terpenoid Transport in Plants: How Far from the Final Picture?

“…After the formation of the precursors, FPP, GPP, and GGPP, the different terpenes are generated through the action of terpene synthases (TPS), which are enzymes implicated in the formation of primary terpene skeletons, and are then further modified by the action of various enzyme classes, such as cytochrome P450 hydroxylases, dehydrogenases, and glycosyl- and methyl-transferases [ 6 , 73 ]. These enzymes have various subcellular localization: for instance, P450 enzymes are generally located in the ER membrane [ 74 ], while other enzymes are placed essentially in the cytosol or in the ER surface [ 75 , 76 , 77 , 78 ]. Differently from terpenes that are synthesized in the cytosol, such as tri- and sesquiterpenes, the biosynthesis of diterpenes and tetraterpenes (and their derivatives) implies the translocation of intermediates from the plastid to the cytosol and/or ER, where the modification enzymes reside.…”

Engineering yeast for the production of plant terpenoids using synthetic biology approaches

“…The isolation of the intra-organelle environment from the rest of the cytoplasm offers a unique advantage for synthetic biology approaches in yeast. 178 The compartmentalization of terpenoid production pathways in organelles provides the ability to sequester a pathway from the endogenous pathways of the cytoplasm 178,179 and has been recently shown to facilitate significant yield improvements. In terms of terpenoid synthesis, organelles would most notably limit the overlap of the heterologous pathways with the endogenous MVA pathway and eliminate the competition for isoprenoid precursors 8 with immediate benefits for target product yield and host growth, respectively.…”

“…170 Differences in organelle structure and metabolic functions confer these compartments a specic cellular environment with unique properties that should be considered for targeting a specic biosynthetic pathway. 1,7 Organelles successfully employed for the production of terpenoids are the mitochondria, peroxisome, and endoplasmic reticulum, 179 recently reviewed by Yocum et al 170 This review will summarize approaches that apply to the production of plant terpenoids in yeast.…”

Engineering the plant metabolic system by exploiting metabolic regulation

“…The optimization of subcellular compartmentalization for PME approaches can include the relocalization of the enzyme production in the organelles, the alteration of the subcellular precursor availability, or the regulation of the transporters associated with biosynthesis (Heinig et al, 2013;Jaramillo-Madrid et al, 2022). For instance, retargeting the expression of the enzymes to the compartment with the highest precursor pool can be used to increase the targeted metabolite accumulation.…”

“…The increased chloroplast size can offer more suitable structures for maximizing metabolic production, such as terpene production (Nagegowda & Gupta, 2020). The ER is also an interesting target organelle to be engineered, since it is the center of the synthesis and transport of a wide diversity of metabolites (Jaramillo‐Madrid et al, 2022). Although currently there are not many studies that have reported modifications of the ER to optimize metabolic production in plants, the studies carried out in yeast showed that ER expansion through knockout of the phosphatidate phosphatase 1 ( ScPAH1 ) gene increases its metabolic capacity (Arendt et al, 2017; Zhao et al, 2021).…”