Lee Jin, Park Hyun-Ah, Shin Kyung-Chul, Oh Deok-Kun
Department of Bioscience and Biotechnology, Konkuk University, 1 Hwayang-dong, Gwangjin-gu, Seoul 143-701, South Korea.
Department of Bioscience and Biotechnology, Hankuk University of Foreign Studies, Mohyein-eup, Cheoin-gu, Yongin-si, Gyeonggi-do 17035, South Korea.
J Biosci Bioeng. 2025 Jan;139(1):1-6. doi: 10.1016/j.jbiosc.2024.09.002. Epub 2024 Oct 15.
Resolvin E series (Rvs), such as RvE4 (5S,15S-dihydroxyeicosapentaenoic acid) and its stereoselective enantiomer (5R,15R-dihydroxyeicosapentaenoic acid), play an important role in promoting the resolution of inflammation and are derived from eicosapentaenoic acid (EPA) by M2 macrophage in human. However, they have been synthesized using expensive and inefficient chemical methods. Here, we performed efficient quantitative production of RvE4 and its enantiomer from EPA using Escherichia coli expressing double-dioxygenating 15S-lipoxygenase (15S-LOX) from Archangium violaceum and double-dioxygenating 15R-LOX from Sorangium cellulosum, respectively, with solvent, polymer, and adsorbent resin. The cell density, substrate concentration, solvent types and concentrations, polymer types and concentrations, and resin concentration were optimized for the enhanced bioconversion of EPA into RvE4 and its enantiomer. Under the optimized conditions, A. violaceum 15S-LOX and S. cellulosum 15R-LOX expressed in E. coli converted 6.0 mM EPA into 4.3 mM (1.44 g/L) RvE4 and 5.8 mM (1.94 g/L) RvE4 enantiomer in 60 min, with productivities of 4.3 and 5.8 mM/h and molar conversions of 72% and 97%, respectively. To date, these are the highest concentrations, productivities, and conversions of RvE4 and its enantiomer. The concentrations of RvE4 and its enantiomer obtained from the conversion of EPA with solvent, polymer, and resin were 2.5- and 3.2-fold higher than those without the additives, respectively.
消退素E系列（Rvs），如RvE4（5S,15S-二羟基二十碳五烯酸）及其立体选择性对映体（5R,15R-二羟基二十碳五烯酸），在促进炎症消退中发挥重要作用，且在人体中由M2巨噬细胞从二十碳五烯酸（EPA）衍生而来。然而，它们一直是通过昂贵且低效的化学方法合成的。在此，我们利用分别表达来自紫色栖热放线菌的双加氧15S-脂氧合酶（15S-LOX）和来自纤维堆囊菌的双加氧15R-脂氧合酶的大肠杆菌，结合溶剂、聚合物和吸附树脂，从EPA高效定量生产RvE4及其对映体。对细胞密度、底物浓度、溶剂类型和浓度、聚合物类型和浓度以及树脂浓度进行了优化，以增强EPA向RvE4及其对映体的生物转化。在优化条件下，大肠杆菌中表达的紫色栖热放线菌15S-LOX和纤维堆囊菌15R-LOX在60分钟内将6.0 mM EPA转化为4.3 mM（1.44 g/L）RvE4和5.8 mM（1.94 g/L）RvE4对映体，生产率分别为4.3和5.8 mM/h，摩尔转化率分别为72%和97%。迄今为止，这些是RvE4及其对映体的最高浓度、生产率和转化率。从EPA与溶剂、聚合物和树脂的转化中获得的RvE4及其对映体的浓度分别比不使用添加剂时高2.5倍和3.2倍。
