Executive Summary : | The growing concern of global warming due to fossil fuel use has sparked a significant interest in developing renewable and eco-friendly biofuels. Alka(e)nes, major components of fossil fuels, are biosynthesised sustainably using fatty acid decarboxylation, a pathway that involves the direct decarboxylation of fatty acids to alka(e)nes by fatty acid decarboxylases. These enzymes produce highly valuable 1-alkenes, which have significant applications in the polymer, lubricant, and detergent industries. Recently, a 1-alkene-producing metalloenzyme, UndB, was discovered in bacteria, which catalyzes the conversion of fatty acids to 1-alkenes. Despite its importance, the enzyme has been poorly characterized due to its association with membranes. This study aims to investigate the membrane-bound metalloenzyme, UndB, responsible for converting fatty acids to terminal alkenes, a chemically challenging reaction. The researchers will conduct purification and biochemical characterization of the enzyme, perform activity, kinetics, mechanism, and reaction intermediates studies, and incorporate the improved enzymes into microorganisms. The ultimate goal is to investigate the efficiency of engineered microorganisms for the biosynthesis of fuel-grade alkenes. The investigation may lead to critical progress in the global efforts of microbial biosynthesis of hydrocarbon-based biofuels. The rigorous investigations on UndB will not only provide fundamental knowledge of 1-alkene production in nature but also direct the development of a highly efficient whole-cell biocatalyst for producing industrially valuable 1-alkenes. |