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Abstract
North Pacific krill (Euphausia pacifica) contain 8R-hydroxy-eicosapentaenoic acid (8R-HEPE), 8R-hydroxy-eicosatetraenoic acid (8R-HETE) and 10R-hydroxy-docosahexaenoic acid (10R-HDHA). These findings indicate that E. pacifica must possess an R type lipoxygenase, although no such enzyme has been identified in krill. We analyzed E. pacifica cDNA sequence using next generation sequencing and identified two lipoxygenase genes (PK-LOX1 and 2). PK-LOX1 and PK-LOX2 encode proteins of 691 and 686 amino acids, respectively. Recombinant PK-LOX1 was generated in Sf9 cells using a baculovirus expression system. PK-LOX1 metabolizes eicosapentaenoic acid (EPA) to 8R-HEPE, arachidonic acid (ARA) to 8R-HETE and docosahexaenoic acid (DHA) to 10R-HDHA. Moreover, PK-LOX1 had higher activity for EPA than ARA and DHA. In addition, PK-LOX1 also metabolizes 17S-HDHA to 10R,17S-dihydroxy-docosahexaenoic acid (10R,17S-DiHDHA). PK-LOX1 is a novel lipoxygenase that acts as an 8R-lipoxygenase for EPA and 10R-lipoxygenase for DHA and 17S-HDHA. Our findings show PK-LOX1 facilitates the enzymatic production of hydroxy fatty acids, which are of value to the healthcare sector.
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Details
1 Iwate Biotechnology Research Center, Kitakami, Japan (GRID:grid.277489.7) (ISNI:0000 0004 0376 441X)
2 Ehime University, Depatment of Bioscience, Graduate School of Agriculture, Matsuyama, Japan (GRID:grid.255464.4) (ISNI:0000 0001 1011 3808)
3 Bizen Chemical Co., Ltd., Akaiwa-shi, Japan (GRID:grid.255464.4)
4 Iwate Biotechnology Research Center, Kitakami, Japan (GRID:grid.277489.7) (ISNI:0000 0004 0376 441X); Teikyo University of Science, Faculty of Life and Environmental Science, Adachi-ku, Japan (GRID:grid.412336.1) (ISNI:0000 0004 1770 1364)