Pentane formation during the anaerobic reactions of reticulocyte lipoxygenase. Comparison with lipoxygenases from soybeans and green pea seeds.

The lipoxygenases from reticulocytes, soybeans and green pea seeds produce pentane in an anaerobic assay containing 13Ls-hydroperoxy-9-cis, 11-trans-octadecadienoic acid and 9,12-all-cis-octadecadienoic acid. The presence of oxygen strongly inhibits pentane formation by the three enzymes. Relative to the lipoxygenase activity with linoleic acid as substrate, soybean lipoxygenase is 4-times as effective in pentane formation as the lipoxygenases from reticulocytes or green pea seeds. Pentane formation by the reticulocyte lipoxygenase is completely inhibited by lipoxygenase inhibitors (5,8,11,14-eicosatetraynoic acid, 3-t-butyl-4-hydroxyanisol) but only partially by radical scavengers which do not influence the oxygenase activity (2,6-di-t-butyl-4-hydroxytoluene). From the temperature dependence below 20 degrees C an activation energy of the pentane production by the reticulocyte lipoxygenase of about 28 kJ/mol was calculated, which is somewhat higher than that for the oxygenase activity. During the anaerobic reaction of both reticulocyte and soybean lipoxygenase C18-oxodienes, C13-oxodienes, linoleic acid dimers and a polar compound proposed to be epoxy-hydroxyoctadecenoic acid are produced in a similar pattern. Reticulocyte lipoxygenase produces pentane with submitochondrial particles only under anaerobic conditions after an aerobic preincubation. During the incubation of intact reticulocytes with the calcium ionophore A23187 or arachidonic acid, pentane is released. Preincubation of the cells with lipoxygenase inhibitors completely abolishes the pentane formation. Erythrocytes do not form any pentane under the same experimental conditions.