Obesity is associated with an increased risk of developing metabolic syndrome, type 2 diabetes and cardiovascular disease. The adipose tissue significantly affects the whole-body metabolism of carbohydrates and lipids, which also involves fatty acid esters of hydroxy fatty acids (FAHFA), lipid mediators that affect glucose uptake and insulin sensitivity in target tissues. Our goal is to identify the metabolic pathways between lipid synthesis and antioxidant defense involved in FAHFA biosynthesis, to examine the substrate specifity of the candidate enzyme, and to introduce a methodology for the quantification of metabolic fluxes through redox reactions linking central carbon metabolism and nicotinamidadene cofactor-driven reductive biosynthesis.