Department of Integrative Bioscience and Biotechnology, Konkuk University, Seoul, Republic of Korea.
School of Biological Sciences and Technology, Chonnam National University, Gwangju, Republic of Korea.
Biotechnol Prog. 2022 Sep;38(5):e3267. doi: 10.1002/btpr.3267. Epub 2022 May 16.
Recombinant Escherichia coli cells expressing 8,11-linoleate diol synthase (LDS) from Penicillium chrysogenum convert oleic and palmitoleic acids to 8-hydroperoxy-9(Z)-octadecenoic acid (HPOME) and 8-hydroperoxy-9(Z)-hexadecenoic acid (HPHME) only, respectively. However, recombinant E. coli cells expressing Q889A variant 6,8-LDS from Penicillium oxalicum as an 8,11-LDS converted oleic and palmitoleic acids to 8,11-dihydroxy-9(Z)-octadecenoic acid (DiHOME) and 8,11-dihydroxy-9(Z)-hexadecenoic acid (DiHHME), respectively, which were identified using liquid chromatography-tandem mass spectrometry analysis. To select suitable variants for producing these compounds, position 889 of 6,8-LDS from P. oxalicum was substituted with other amino acids, and recombinant E. coli cells expressing Q889L and Q889A variants were chosen as the best biocatalysts for producing 8,11-DiHOME and 8,11-DiHHME, respectively. The optimal conditions for producing 8,11-DiHOME or 8,11-DiHHME using cells expressing Q889L or Q889A variant 6,8-LDS were pH 6.5 and 30 °C with 5% (v/v) dimethyl sulfoxide, 60 g L cells, and 10 g L oleic acid or 7.5 g L palmitoleic acid, respectively. Under these conditions, 10.7 g L 8,11-DiHOME and 8.1 g L 8,11-DiHHME were produced for 1.5 h with molar yields of 96.4% and 96.2% and productivities of 7.1 and 5.4 g L h , respectively. The molar yields and concentrations of 8,11-DiHOME and 8,11-DiHHME were highest among those of other reported DiHOMEs and DiHHMEs. To the best of our knowledge, this is the first quantitative production of 8,11-DiHOME and 8,11-DiHHME.
从产黄青霉（Penicillium chrysogenum）中表达的 8,11-亚油酸二醇合酶（LDS）的重组大肠杆菌细胞仅将油酸和棕榈油酸分别转化为 8-羟基-9(Z)-十八碳烯酸（HPOME）和 8-羟基-9(Z)-十六碳烯酸（HPHME）。然而，从构巢曲霉（Penicillium oxalicum）中表达的 Q889A 变体 6,8-LDS 的重组大肠杆菌细胞将油酸和棕榈油酸分别转化为 8,11-二羟基-9(Z)-十八碳烯酸（DiHOME）和 8,11-二羟基-9(Z)-十六碳烯酸（DiHHME），这两种物质均通过液相色谱-串联质谱分析鉴定。为了选择合适的变体来生产这些化合物，构巢曲霉 6,8-LDS 的 889 位氨基酸被替换为其他氨基酸，表达 Q889L 和 Q889A 变体的重组大肠杆菌细胞分别被选为生产 8,11-DiHOME 和 8,11-DiHHME 的最佳生物催化剂。用表达 Q889L 或 Q889A 变体 6,8-LDS 的细胞生产 8,11-DiHOME 或 8,11-DiHHME 的最佳条件分别为 pH 6.5、30°C 和 5%（v/v）二甲基亚砜、60 g L 细胞和 10 g L 油酸或 7.5 g L 棕榈油酸。在这些条件下，1.5 h 内可分别产生 10.7 g L 8,11-DiHOME 和 8.1 g L 8,11-DiHHME，摩尔收率分别为 96.4%和 96.2%，产率分别为 7.1 和 5.4 g L h 。8,11-DiHOME 和 8,11-DiHHME 的摩尔产率和浓度在其他报道的 DiHOMEs 和 DiHHMEs 中最高。据我们所知，这是首次定量生产 8,11-DiHOME 和 8,11-DiHHME。
