Investigating The Multiple Enzyme Complex In Carotenoid Biosynthesis Pathway

In nature, plant carotenoids are 40-carbon isoprenoids with polyene chains that may have double bonds. They are vital components of all photosynthetic organisms, playing several roles in the plant cell, including photosynthesis, pigmentation, phytohormone synthesis, signaling and photoprotection. In this study, we investigated protein-protein interactions among key enzymes involved in the carotenoid biosynthesis pathway using the Bimolecular Fluorescent Complementation (BiFC) assay. Our results revealed several interactions that suggest a coordinated regulation of enzymatic activity within this pathway, highlighting potential regulatory complexes that influence carotenoid production. The study also highlights the pivotal role of Zeta-Carotene Isomerase (Z-ISO) as a key regulatory enzyme in carotenoid biosynthesis. Transgenic Arabidopsis thaliana lines expressing YFP-tagged constructs were successfully generated and exhibited detectable fluorescence, providing essential tools for future in vivo analyses of enzyme localization and interaction dynamics. These findings enhance our understanding of carotenoid biosynthesis regulation and offer tools for further functional and mechanistic studies.