Department of Bioscience and Biotechnology, Konkuk University, 120 Neungdong-ro, Gwangjin-gu, Seoul, 05029, Republic of Korea.
Forest Plant Industry Department, Baekdudaegan National Arboretum, Bonghwa, 36209, Republic of Korea.
Biotechnol Lett. 2019 May;41(4-5):575-582. doi: 10.1007/s10529-019-02659-5. Epub 2019 Mar 2.
To quantitatively hydroxylate 8S- and 10S-positions on polyunsaturated fatty acids by recombinant Escherichia coli cells expressing mouse arachidonate 8S-lipoxygenase (8S-LOX).
Hydroxylated products gained from the conversion of arachidonic acid (20:4, AA), eicosapentanoic acid (20:5, EPA), and (22:6, DHA) by recombinant E. coli cells containing 8S-LOX from mouse were identified as 8S-hydroxy-5,9,11,14(Z,E,Z,Z)-eicosatetranoic acid (8S-HETE), 8S-hydroxy-5,9,11,14,17(Z,E,Z,Z,Z)-eicosapentanoic acid (8S-HEPE), and 10S-hydroxy-4,8,12,14,16,19(Z,E,Z,Z,Z,Z)-docosahexaenoic acid (10S-HDoHE), respectively. Under the optimal hydroxylation conditions of pH 7.5, 30 °C, 5% (v/v) ethanol, 15 g cells l, and 5 mM substrate, AA, EPA, and DHA were hydroxylated into 4.37 mM 8S-HETE, 3.77 mM 8S-HEPE, and 3.13 mM 10S-HDoHE for 60, 90, and 60 min, with 87, 75, and 63% molar conversions, respectively.
To the best of our knowledge, this is the first quantitatively biotechnological production of 8S-HETE, 8S-HEPE, and 10S-HDoHE.
通过表达鼠花生四烯酸 8S-脂氧合酶（8S-LOX）的重组大肠杆菌细胞定量羟基化多不饱和脂肪酸的 8S-和 10S-位。
通过含有来自鼠的 8S-LOX 的重组大肠杆菌细胞转化花生四烯酸（20:4，AA）、二十碳五烯酸（20:5，EPA）和二十二碳六烯酸（22:6，DHA）获得的羟基化产物被鉴定为 8S-羟基-5,9,11,14(Z,E,Z,Z)-二十碳四烯酸（8S-HETE）、8S-羟基-5,9,11,14,17(Z,E,Z,Z,Z)-二十碳五烯酸（8S-HEPE）和 10S-羟基-4,8,12,14,16,19(Z,E,Z,Z,Z,Z)-二十二碳六烯酸（10S-HDoHE）。在最佳羟基化条件（pH 7.5、30°C、5%（v/v）乙醇、15 g 细胞 l 和 5 mM 底物）下，AA、EPA 和 DHA 分别在 60、90 和 60 min 内羟基化为 4.37 mM 8S-HETE、3.77 mM 8S-HEPE 和 3.13 mM 10S-HDoHE，摩尔转化率分别为 87%、75%和 63%。
据我们所知，这是首次定量生物技术生产 8S-HETE、8S-HEPE 和 10S-HDoHE。
