Figure 5

Time course of turnover of 13-HOD by glyoxysomes from etiolated sunflower. β-oxidation assays were prepared as described in the material and method section. HPLC-traces after different reaction times were detected at 260 nm (A) and 234 nm (B). Absorption signals of the individual intermediates are indicated by open (234 nm, conjugated double bond) or closed (260 nm, CoA-ester) circles and open squares (280 nm, keto-diene system). Besides 13-HOD-CoA (retention time ~ 101 min) acyl-CoAs bearing the conjugated hydroxy-diene system after one (retention time ~ 97 min) and two (retention time ~ 93 min) cycles of β-oxidation are detected and have been validated by MS. The respective oxidized KOD-derivatives elute with slightly elongated retention times (~ 102, 98 and 94 min). Intermediates lacking the 234 nm absorption coelute with authentic acyl-CoA standards with chain length between 4 and 12 carbons.