Partial Purification and Characterization of UDP-Glucose Pyrophosphorylase from Immature Grains of Wheat (Triticum aestivum L.)

Uridine diphosphate glucose pyrophosphorylase (UDP-Glc PPase, EC2.7.7.9) was purified 65 fold from immature grains of wheat (Triticum aestivum L. cv, WH-147) by ammonium sulphate fractionation, DEAE-cellulose anion exchange chromatography and Sephadex G-100 permeation chromatography. The partially purified enzyme, having molecular weight of 72 kD, exhibited broad pH optimum between 8 and 9 and was stable at 4°C for 15 days. At pH 8.5, the enzyme followed typical hyperbolic kinetics with respect to UDP-glucose and inorganic pyrophosphate (Km 0.22 mM and 0.66 mM respectively). The enzyme showed absolute requirement for Mg²⁺ and did not appear to require sulfhydryl groups for its activity. Initial velocity and product inhibition studies indicated sequential addition of substrates and sequential release of products.