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A. B. 200. 140. Linoleic acid. Linoleic acid. Arachidonic acid. 120. 150. 100. 80. Lipoxygenase activity (nkat mg -1 ). Lipoxygenase activity (nkat mg -1 ). 100. 60. 40. 50. 20. 0. 0. 4. 5. 6. 7. 8. 9. 10. 20. 30. pH. Reaction Time (min). C. 250.
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A B 200 140 Linoleic acid Linoleic acid Arachidonic acid 120 150 100 80 Lipoxygenase activity (nkat mg-1) Lipoxygenase activity (nkat mg-1) 100 60 40 50 20 0 0 4 5 6 7 8 9 10 20 30 pH Reaction Time (min) C 250 Linoleic acid 5 μM 200 15 μM 30 μM 60 μM 150 Lipoxygenase activity (nkat mg-1) 100 μM 150 μM 100 200 μM 50 0 5 10 15 20 25 30 Reaction time (min) Supplemental Figure S3. LOX activity of recombinant His-tag-CaLOX1 expressed in E. coli BL21 (DE3). A, Effect of pH on CaLOX1 LOX activity. Purified CaLOX1 was analyzed for enzyme activity in phosphate buffers with different pH values using 0.1 mM linoleic acid as a substrate. B, Time courses of CaLOX1 LOX activity. Purified CaLOX1 was incubated with 0.1 mM of linoleic acid or arachidonic acid as a substrate. C, Time courses of the LOX activity of purified CaLOX1, as determined with various concentrations of linoleic acid as a substrate.