In this study, the conversions of 1-dodecanoic, ω-hydroxydodecanoic acid and α,ω-dodecanedioic acid using whole cell biotransformation of Escherichia coli BW25113ΔfadD expressing CAR and ADH enzymes were demonstrated. First 13 CAR enzymes were examined for 1-dodecanoic acid reduction, and CAR encoded by mab4714 from Mycobacterium abscessus showed the highest conversion of 53.1% in single cells of heterologous CAR and endogenous ADH. For a better conversion, the host cells were engineered to simultaneously express Yarrowia lipolytica ADH2 with the GroES/EL-DnaK/J/E chaperone in a single host system. In addition, two-cell system using two strains of E. coli expressing CAR-Sfp and ADH-GroES/EL-DnaK/J/E was also investigated. In results, additional ADH expression was not effective in a single host system, whereas two cell system significantly increased α,ω-dodecanedioic acid conversion by total 71.3%; α,ω-dodecanediol (68.2%) and ω-hydroxydodecanoic acid (3.1%), respectively. Interestingly, the MAB4714 CAR enzyme could converted ω-hydroxydodecanoic acid into α,ω-dodecanediol up to 97.2% conversion in 17 h (12.4 mg/L/h). Finally, structural understanding of the higher activity against ω-hydroxydodecanoic was understood by docking simulations which suggested hydrogen-bonding interactions between ω-hydroxyl group and polar residues such as Gln434 and Thr285 were holding the substrate tightly with more stable positioning in the active site.

在这项研究中，使用的全细胞生物转化1-十二烷，ω羟基十二烷酸和α，ω-十二烷二酸的转化大肠杆菌BW25113 Δ的fadD表达CAR和ADH酶进行了论证。检查了前 13 种 CAR 酶的 1-十二烷酸还原，并且由来自脓肿分枝杆菌的mab4714编码的 CAR在异源 CAR 和内源性 ADH 的单细胞中显示出最高的转化率为 53.1%。为了更好地转化，宿主细胞被设计为在单个宿主系统中同时表达解脂耶氏酵母ADH2 和 GroES/EL-DnaK/J/E 伴侣。此外，使用两种大肠杆菌菌株的双细胞系统还研究了表达 CAR-Sfp 和 ADH-GroES/EL-DnaK/J/E。结果，额外的 ADH 表达在单个宿主系统中无效，而两个细胞系统显着增加了 α,ω-十二烷二酸转化率，总计 71.3%；分别为 α,ω-十二烷二醇 (68.2%) 和 ω-羟基十二烷酸 (3.1%)。有趣的是，MAB4714 CAR 酶可以在 17 小时内将 ω-羟基十二烷酸转化为 α,ω-十二烷二醇，转化率高达 97.2%（12.4 mg/L/h）。最后，通过对接模拟了解了对 ω-羟基十二烷酸的更高活性的结构理解，这表明 ω-羟基与极性残基（如 Gln434 和 Thr285）之间的氢键相互作用将底物紧紧地固定在活性位点中，并具有更稳定的定位。