The EThe Effect of Flavonoids on Pyruvate Carboxylase Enzyme Activity: An In Vitro and In Silico Approachffect of Flavonoids on Pyruvate Carboxylase Enzyme Activity: An In Vitro and In Silico Approach

Abstract

Pyruvate carboxylase (PC), the enzyme responsible for catalyzing oxaloacetate synthesis, serves as a central regulator of gluconeogenesis and lipogenesis, with its hyperactivity playing a fundamental role in the pathogenesis of type 2 diabetes. This study aims to quantitatively and thermodynamically evaluate the modulatory effects of polyphenol-derived bioactive compounds—specifically flavonoids like quercetin and rutin—on the catalytic activity of the PC enzyme. By integrating in vitro spectrophotometric assays with in silico molecular docking techniques, the binding kinetics of flavonoids to the enzyme's allosteric domain were thoroughly investigated. Empirical data obtained within the scope of the research proved that quercetin and its derivatives competitively inhibit the ATP-dependent carboxylation phase. During the formation of the substrate-enzyme complex, the quercetin molecule was noted to significantly reduce catalytic efficiency. The dynamics of the obtained results provide a molecular foundation for the development of next-generation pharmacotherapeutic agents based on plant-derived flavonoids, designed to correct carbohydrate metabolism disorders and block excessive glucose production in hepatocytes.