FIG 3.

Peroxygenase activities of the purified recombinant AfPXG. (A) Co-oxidation of radiolabeled polyunsaturated fatty acids in the presence of cumene hydroperoxide (CuOOH), hydrogen peroxide (H₂O₂), 9-hydroperoxy-10,12-octadecadienoic acid (9-HPOD), or 13-hydroperoxy-9,11-octadecadienoic acid (13-HPOD). ¹⁴C-labeled substrates metabolized by purified recombinant AfPXG were separated by TLC and analyzed by radiodetection. C_18:1, oleic acid; C_18:2, linoleic acid; C_18:3, linolenic acid. *, P < 0.05; **, P < 0.01; ***, P < 0.001. Different lowercase letters indicate significant differences (P < 0.05) for the fatty acids used. (B and C) UV-HPLC analysis of the metabolization of fatty acid hydroperoxides by purified AfPXG. (I) Products formed after 2 h of incubation at 27°C of 13-HPOD or 9-HPOD by AfPXG; (II) 13-HOD or 9-HOD standards; (III) 13-HPOD or 13-HPOD incubated with heat-inactivated AfPXG. 9-HPOD and 13-HPOD did not differ in their ability to co-oxidize the reactions when the difference was analyzed by the t test. This FAOOH reduction capacity was significantly different from that measured in the presence of H₂O₂ and cumene hydroperoxide when analyzed by the t test.