Abstract A robust Escherichia coli (E. coli) strain, with a dual protection system to fight against microbial contamination and T7 phage infection, is used as a chassis to overexpress various proteins in auxotrophic MOPS medium. Among them, a robust EH (epoxide hydrolase)-producing E. coli was used as a biocatalyst to enantioselectively resolve racemic glycidyl o-methylphenyl ether (rac-o-GMPE) for preparing (R)-o-GMPE with 6416.7 U/g wet cells. The optimal temperature, pH and biocatalyst dosage were 25 °C, 7.0 and 0.2 mg/mL in aqueous buffer, respectively. The maximal yield (23.0 %) and ee (99.2 %) were achieved within 12 min in 20 mM rac-o-GMPE. Additionally, 98.0 % ee and 38.7 % yield were observed by cell-supplemental strategy with substrate concentration of 100 mM. Furthermore, to alleviate substrate inhibition and product toxicity toward the cells, a biphasic system (aqueous buffer: butyl acetate = 0.5:1) was developed to catalyze 3 M rac-o-GMPE with the initial reaction rate of 549.7 mmol/L/h, 91.7 % ee, 17.1 % yield and 3.66 g/L/h space-time yield.

中文翻译：

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在强健大肠杆菌菌株中的高效蛋白质表达及其在高浓度外消旋缩水甘油邻甲基苯基醚的动力学拆分中的应用

摘要 一种具有双重保护系统以对抗微生物污染和 T7 噬菌体感染的强健大肠杆菌 (E.coli) 菌株被用作底盘在营养缺陷型 MOPS 培养基中过表达各种蛋白质。其中，一种强大的 EH（环氧化物水解酶）生产大肠杆菌被用作生物催化剂，对映选择性拆分外消旋缩水甘油基邻甲基苯基醚（rac-o-GMPE）以制备（R）-o-GMPE，6416.7 U/g湿细胞。在水性缓冲液中，最佳温度、pH 值和生物催化剂用量分别为 25 °C、7.0 和 0.2 mg/mL。在 20 mM rac-o-GMPE 中，在 12 分钟内达到最大产率 (23.0%) 和 ee (99.2%)。此外，通过底物浓度为 100 mM 的细胞补充策略观察到 98.0% ee 和 38.7% 产率。此外，