Characterization of a Selenium-Independent Glutathione Peroxidase From Euglena gracilis.

Light or dark grown Euglena gracilis strains contain similar levels of glutathione (GSH) peroxidase. Cells in midstationary phase of growth contained the highest level of the enzyme. The enzyme was purified 280-fold to homogeneity from the permanently bleached strain, E. gracilis var bacillaris W(3)BUL. The native enzyme has a molecular weight of 130,000 as measured by gel permeation chromatography, and contains four subunits (mol wt 31,500) as measured by sodium dodecyl sulfate gel electrophoresis. A variable amount of a higher molecular weight form of the enzyme (approximate mol wt 250,000) was detected but not further characterized. The enzyme has an isoelectric point of 4.7. No selenium could be detected in the purified enzyme. The enzyme is active with H(2)O(2) and a variety of organic hydroperoxides, including 13-hydroperoxylinoleic acid, and is specific for GSH as the thiol substrate. Apparent K(m) values for H(2)O(2), t-butyl hydroperoxide, and GSH were 0.03, 1.5, and 0.7 millimolar, respectively. A comparison of selenium-dependent and selenium-independent GSH peroxidases from various eukaryotic sources is presented.