Figure 2.

NP-HPLC analysis of three products generated from DHA by Osc-LOX enzymatic catalysis. The reactions were carried out using 50 mM Tris–HCl buffer (pH 8.0), varying amounts of enzyme 50–400 Units mL⁻¹, and substrate 50 μM for 30 min at 30 °C. 25 mM sodium borohydride was used for deoxygenation of hydro peroxide products after enzyme reaction. (a) DHA standard eluted at 2.65 min. (b) 17S-HDHA standard eluted at 5.57 min. (c) Resolvin D5 standard eluted at 12.3 min. (d) First product (17S-HDHA) was generated from DHA by Osc-LOX concentration of 50 Unit mL⁻¹. (e) Second product (RvD5) was generated from 17S-HDHA by Osc-LOX concentration of 100 Unit mL⁻¹ but not completely converted. (f) When using Osc-LOX concentration of 200 Unit mL⁻¹, 17S-HDHA was completely converted into RvD5. (g) Final product was generated from RvD5 by Osc-LOX concentration of 400 Unit mL⁻¹.