ABSTRACT Plant metabolism is increasingly being demonstrated to be partially controlled by dynamically assembled metabolons—multienzyme complexes that enable substrate channeling, insulate reactive intermediates, and permit rapid, low‐energy flux control. Rigorous criteria are defined to distinguish true metabolons from generic assemblies, and evidence is synthesized across cyanogenic glucoside, phenylpropanoid/flavonoid, alkaloid, terpenoid, polyamine, sporopollenin, and auxin pathways. A practical workflow is presented in which AP‐MS (Affinity purification mass spectrometry)/Co‐IP (Co‐immunoprecipitation), proximity labeling, BiFC (Bimolecular fluorescence complementation)/FRET (Förster resonance energy transfer)/Split‐luciferase, and isotope‐dilution metabolomics are integrated to resolve composition, dynamics, and direct channeling in vivo . In enzyme‐based substrate channeling engineering, design rules are distilled for membrane anchoring, modular scaffolds, compartment targeting, and inducible/optogenetic control, and limitations such as metabolic burden, stoichiometry, and leakiness are noted. An AI‐assisted loop is outlined in which structure‐aware generative models produce binders/interfaces that are coupled to spatial optimization of enzyme order, orientation, and distance. Together, these advances reposition metabolons as a deployable technology for programmable flux in plants, enabling safer handling of labile intermediates and higher titers of valuable natural products.
Zhang et al. (Mon,) studied this question.