Purification and Substrate Specificity of Key Enzymes Involved in Plant Wax Synthesis

Plant waxes, as a class of hydrophobic lipid complexes covering the outer surface of plant epidermal cells, play a central role in reducing water transpiration, resisting biotic and abiotic stresses, and maintaining epidermal integrity. The biosynthesis of plant waxes is a complex multi-enzyme catalyzed process, in which ketolipid acyl-CoA synthase (KCS), fatty acyl-CoA reductase (FAR), and wax synthase (WSD) are key enzymes regulating the synthesis of its core components. The catalytic activity and substrate specificity of these three enzymes directly determine the composition, content, and physiological functions of waxes. However, KCS, FAR, and WSD currently suffer from issues such as low expression levels in vivo, poor natural stability, and difficulties in purification. Based on this, this paper uses Arabidopsis thaliana and Brassica napus epidermal tissues as materials to optimize purification process parameters and verify the applicability of the system. Simultaneously, it systematically analyzes the substrate specificity patterns of the three enzymes, providing core technical support and theoretical basis for further analyzing the regulatory mechanisms of plant wax synthesis.