Characterization of Arachidonate 5S-Lipoxygenase from Danio rerio with High Activity for the Production of 5S- and 7S-Hydroxy Polyunsaturated Fatty Acids

Biotransformation of Polyunsaturated Fatty Acids to Trioxilins by Lipoxygenase from Pleurotus sajor‐caju

“…Through gel‐filtration chromatography, the total molecular mass of the native enzyme was estimated to be 146 kDa, indicating that the enzyme exists as a homodimer (Figure 1B). This dimeric form is consistent with most LOXs, including 15 S ‐LOXs from Burkholderia thailandensis and rabbit, [25,26] 12 S ‐LOXs from Endozoicomonas numazuensis , Myxococcus xanthus , and human, [27–29] and 5 S ‐LOXs from Danio rerio and human [29,30] . A LOX with 98 % sequence identity to this enzyme had been previously reported, [31] but its association form has not been identified.…”

“…This dimeric form is consistent with most LOXs, including 15S-LOXs from Burkholderia thailandensis and rabbit, [25,26] 12S-LOXs from Endozoicomonas numazuensis, Myxococcus xanthus, and human, [27][28][29] and 5S-LOXs from Danio rerio and human. [29,30] A LOX with 98 % sequence identity to this enzyme had been previously reported, [31] but its association form has not been identified.…”

Biotransformation of C20- and C22-polyunsaturated fatty acids and fish oil hydrolyzates to R,R-dihydroxy fatty acids as lipid mediators using double-oxygenating 15R-lipoxygenase

“…5 R ,15 R -Dihydroxy-6 E ,8 Z ,11 Z ,13 E -eicosatetraenoic acid (compound 4 ; 5 R ,15 R -DiHETE), 5 R ,15 S -, and 5 S ,15 R -DiHETEs were biosynthesized from 5 R -, 15 R -, and 5 R -HETE standards by 15 R -LOX from S. cellulosum , 15 S -LOX from Burkholderia thailandensis , 29 and 5 S -LOX from Danio rerio . 30 The reactions were performed at 25 °C in a 500 mL baffled flask containing 100 mL of reaction solution in 50 mM 2-[4-(2-hydroxyethyl)piperazin-1-yl]ethanesulfonic acid (HEPES) buffer (pH 8.0) that contained 0.1 mM 15 R - or 5 R -HETE standard, 0.05 mg mL −1 purified enzyme, and 100 mM l -cysteine as a reducing agent for 20 min. The reactions to produce MonoHFAs and DiHFAs, namely 3 , 15 R -hydroxy-5 Z ,8 Z ,11 Z ,13 E ,17 Z -eicosapentaenoic acid (compound 7 ; 15 R -HEPA), and 17 R -hydroxy-4 Z ,7 Z ,10 Z ,13 Z ,15 E ,19 Z -docosahexaenoic acid (compound 11 ; 17 R -HDHA); and 5 R ,15 R -dihydroxy-6 E ,8 Z ,11 Z ,13 E ,17 Z -eicosapentaenoic acid (compound 8 ; 5 R ,15 R -DiHEPA) and 7 R ,17 R -dihydroxy-4 Z ,8 E ,10 Z ,13 Z ,15 E ,19 Z -docosahexaenoic acid (compound 12 ; 7 R ,17 R -DiHDHA) using 15 R -LOX from S. cellulosum were performed at 25 °C in the flask in 50 mM HEPES buffer (pH 8.0) containing 2.0 mM PUFAs ( 1 , 5 , or 9 ), 0.5 or 1.0 g L −1 of cells, and 100 mM l -cysteine for 30 or 60 min, respectively.…”