Department of Bioscience and Biotechnology, Konkuk University, 120 Neungdong-ro, Gwangjin-gu, Seoul, 05029, Republic of Korea.
Department of Integrative Bioscience and Biotechnology, Konkuk University, 120 Neungdong-ro, Gwangjin-gu, Seoul, 05029, Republic of Korea.
Biotechnol Lett. 2022 Sep;44(9):1027-1036. doi: 10.1007/s10529-022-03278-3. Epub 2022 Jul 14.
Peroxidation and reduction of 11S- and 13S-positions on C20 and C22 polyunsaturated fatty acids (PUFAs) by Escherichia coli expressing highly active arachidonate (ARA) 11S-lipoxygenase (11S-LOX) from Enhygromyxa salina with the reducing agent cysteine.
The specific activity and catalytic efficiency of ARA 11S-LOX from E. salina were 4.1- and 91-fold higher than those of only reported ARA 11S-LOX from Myxococcus xanthus, respectively. The hydroxy fatty acids (HFAs) obtained by the biotransformation of ARA, eicosapentaenoic acid (EPA), docosapentaenoic acid (DPA), and docosahexanoic acid (DHA) by Escherichia coli expressing 11S-LOX from E. salina in the presence of cysteine were identified as 11S-hydroxyeicosatetraenoic acid (11S-HETE), 11S-hydroxyeicosapentaenoic acid (11S-HEPE), 13S-hydroxydocosapentaenoic acid (13S-HDPA), and 13S-hydroxydocosahexaenoic acid (13S-HDHA), respectively. The recombinant cells converted 3 mM of ARA, EPA, DPA, and DHA into 2.9 mM of 11S-HETE, 2.4 mM 11S-HEPE, 1. 9 mM 13S-HDPA, and 2.2 mM 13S-HDHA in 60, 80, 120, and 120 min, corresponding to productivities of 72.5, 40.4, 18.5, and 22.4 μM min and conversion yields of 96.7, 80.0, 62.3, and 74.6%, respectively.
We report the highest concentrations, conversion yields, and productivities of 11S- and 13S-hydroxy fatty acids from C20- and C22-PUFAs achieved via E. coli expressing highly active E. salina 11S-LOX.
通过表达来自盐生埃默森小球藻（Enhygromyxa salina）的高活性花生四烯酸（ARA）11S-脂氧合酶（11S-LOX）的大肠杆菌，用还原剂半胱氨酸对 C20 和 C22 多不饱和脂肪酸（PUFAs）的 11S-和 13S-位进行氧化还原反应。
盐生埃默森小球藻的 ARA 11S-LOX 的比活性和催化效率分别比仅报道的粘球菌（Myxococcus xanthus）的 ARA 11S-LOX 高 4.1 倍和 91 倍。大肠杆菌在半胱氨酸存在下表达 11S-LOX 对 ARA、二十碳五烯酸（EPA）、二十二碳五烯酸（DPA）和二十二碳六烯酸（DHA）进行生物转化得到的羟基脂肪酸（HFAs）分别鉴定为 11S-羟基二十碳四烯酸（11S-HETE）、11S-羟基二十碳五烯酸（11S-HEPE）、13S-羟基二十二碳五烯酸（13S-HDPA）和 13S-羟基二十二碳六烯酸（13S-HDHA）。重组细胞在 60、80、120 和 120 min 内将 3 mM 的 ARA、EPA、DPA 和 DHA 转化为 2.9 mM 的 11S-HETE、2.4 mM 的 11S-HEPE、1.9 mM 的 13S-HDPA 和 2.2 mM 的 13S-HDHA，对应的产物生成率为 72.5、40.4、18.5 和 22.4 μM·min 和转化率为 96.7、80.0、62.3 和 74.6%。
我们报告了通过表达来自盐生埃默森小球藻的高活性 11S-LOX 的大肠杆菌从 C20-和 C22-PUFAs 中获得的 11S-和 13S-羟基脂肪酸的最高浓度、转化率和产物生成率。
