Department of Bioscience and Biotechnology, Konkuk University, Seoul 05029, South Korea.
Department of Bioscience and Biotechnology, Konkuk University, Seoul 05029, South Korea
J Lipid Res. 2018 Nov;59(11):2153-2163. doi: 10.1194/jlr.M088823. Epub 2018 Sep 26.
Lipoxygenases (LOXs) catalyze the dioxygenation of PUFAs to produce regio- and stereospecific oxygenated fatty acids. The identification of regio- and stereospecific LOXs is important because their specific products are involved in different physiological activities in various organisms. Bacterial LOXs are found only in some proteobacteria and cyanobacteria, and they are not stable in vitro. Here, we used C20 and C22 PUFAs such as arachidonic acid (ARA), eicosapentaenoic acid, and docosahexaenoic acid to identify an 11-specific LOX from the proteobacterium and explore its in vitro stability and activity. The activity and stability of ARA 11-LOX as well as the production of 11-hydroxyeicosatetraenoic acid from ARA were significantly increased by the addition of phosphatidylcholine, Ca, and coactosin-like protein (newly identified in the yeast ) as stimulatory factors; in fact, LOX activity in the presence of all three factors increased approximately 3-fold. Our results indicate that these stimulatory factors can be used to increase the activity and stability of bacterial LOX and the production of bioactive hydroxy fatty acids, which can contribute to new academic research.
脂氧合酶（LOXs）催化多不饱和脂肪酸（PUFAs）的双加氧作用，生成区域和立体特异性氧合脂肪酸。鉴定区域和立体特异性 LOXs 很重要，因为它们的特定产物参与了不同生物体中的不同生理活动。细菌 LOXs 仅存在于某些 Proteobacteria 和 Cyanobacteria 中，并且在体外不稳定。在这里，我们使用 C20 和 C22 PUFAs，如花生四烯酸（ARA）、二十碳五烯酸（EPA）和二十二碳六烯酸（DHA），从 Proteobacteria 中鉴定出一种 11-特异性 LOX，并探索其体外稳定性和活性。添加磷脂酰胆碱、Ca 和 coactosin 样蛋白（在酵母中鉴定的新蛋白）作为刺激因子可显著增加 ARA 11-LOX 的活性和稳定性，以及 ARA 生成 11-羟二十碳四烯酸；事实上，在这三种因子存在的情况下，LOX 活性增加了约 3 倍。我们的结果表明，这些刺激因子可用于提高细菌 LOX 的活性和稳定性以及生物活性羟脂肪酸的生成，这有助于新的学术研究。
