PARTIAL PURIFICATION AND PROPERTIES OF PUMPKIN LIPOXYGENASE WITH CAROTENE-BLEACHING ACTIVITY

From locally grown pumpkins, an active lipoxygenase preparation with an active carotene-bleaching factor was partially purified by ammonium sulfate fractionation, gel filtration, and ion-exchange chromatography. The enzyme had a pH optimum at 6.5 and was inactive at pH below 3 and above 10. The maximum activity occurred at 30°C. The apparent K_m determined in the presence of linoleate was 0.33 × 10^?3 M. The heavy metals Hg²⁺, Cu²⁺, Co²⁺, and Fe³⁺ were effective inhibitors of this enzyme. Cyanide, fluoride, and L-ascorbic acid also inhibited lipoxygenase activity. Carotene-bleaching activities operated strongly on the lipoxygenase fraction and slightly on the denatured lipoxygenase and hemoproteins treated with heat.