LIPOXYGENASE ACTIVITY ASSOCIATED WITH CYANIDE-INSENSITIVE OXYGEN UPTAKE IN MITOCHONDRIAL FRACTIONS FROM SEEDLINGS OF GLYCINE MAX L

Abstract

Soybean seeds are known to contain high levels of lipoxygenase activity, especially during early stages of germination. Crude mitochondrial fractions from germinating soybeans also have been shown to exhibit high rates of cyanide-insensitive oxygen uptake. These results show the effects of successive discontinuous PercollR density gradients on mitochondrial fractions from 2 day old soybean seeds as judged by polarographic studies and Ouchterlony double diffusion. Axis mitochondria exhibited totally cyanide-sensitive oxygen uptake after two gradients and cotyledon mitochondria exhibited from none to 11% cyanide-insensitive oxygen uptake after three gradients. Mitochondrial fractions which were assayed for lipoxygenase with double diffusion exhibited positive results with fractions that showed cyanide-insensitive oxygen uptake and negative results with cyanide-sensitive mitochondria. These results suggest that lipoxygenase can loosely associate with the mitochondrial membrane and that gradient centrifugation can purify mitochondria free of both lipoxygenase and cyanide-insensitive oxygen uptake.