Key Laboratory of Pharmaceutical Engineering of Zhejiang Province, College of Pharmaceutical Science, Zhejiang University of Technology, Hangzhou, 310014, People's Republic of China.
Key Laboratory for Green Pharmaceutical Technologies and Related Equipment of Ministry of Education, Zhejiang University of Technology, Hangzhou, 310014, People's Republic of China.
Appl Biochem Biotechnol. 2024 Mar;196(3):1509-1526. doi: 10.1007/s12010-023-04641-4. Epub 2023 Jul 10.
Chiral trifluoromethyl alcohols as vital intermediates are of great interest in fine chemicals and especially in pharmaceutical synthesis. In this work, a novel isolate Kosakonia radicincitans ZJPH202011 was firstly employed as biocatalyst for the synthesis of (R)-1-(4-bromophenyl)-2,2,2-trifluoroethanol ((R)-BPFL) with good enantioselectivity. By optimizing fermentation conditions and bioreduction parameters in aqueous buffer system, the substrate concentration of 1-(4-bromophenyl)-2,2,2-trifluoroethanone (BPFO) was doubled from 10 to 20 mM, and the enantiomeric excess (ee) value for (R)-BPFL increased from 88.8 to 96.4%. To improve biocatalytic efficiency by strengthening the mass-transfer rate, natural deep-eutectic solvents, surfactants and cyclodextrins (CDs) were introduced separately in the reaction system as cosolvent. Among them, L-carnitine: lysine (C: Lys, molar ratio 1:2), Tween 20 and γ-CD manifested higher (R)-BPFL yield compared with other same kind of cosolvents. Furthermore, based on the excellent performance of both Tween 20 and C: Lys (1:2) in enhancing BPFO solubility and ameliorating cell permeability, a Tween 20/C: Lys (1:2)-containing integrated reaction system was then established for efficient bioproduction of (R)-BPFL. After optimizing the critical factors involved in BPFO bioreduction in this synergistic reaction system, BPFO loading increased up to 45 mM and the yield reached 90.0% within 9 h, comparatively only 37.6% yield was acquired in neat aqueous buffer. This is the first report on K. radicincitans cells as new biocatalyst applied in (R)-BPFL preparation, and the developed Tween 20/C: Lys-containing synergistic reaction system has great potential for the synthesis of various chiral alcohols.
手性三氟甲基醇作为重要的中间体,在精细化学品中,特别是在药物合成中具有重要的应用价值。在这项工作中,我们首次将新型分离菌Kosakonia radicincitans ZJPH202011 用作生物催化剂,用于合成(R)-1-(4-溴苯基)-2,2,2-三氟乙醇((R)-BPFL),具有良好的对映选择性。通过优化发酵条件和水相缓冲体系中的生物还原参数,将 1-(4-溴苯基)-2,2,2-三氟乙酮(BPFO)的底物浓度从 10mM 提高到 20mM,(R)-BPFL 的对映体过量(ee)值从 88.8%提高到 96.4%。为了通过强化传质速率来提高生物催化效率,我们分别在反应体系中引入天然深共熔溶剂、表面活性剂和环糊精(CDs)作为共溶剂。其中,L-肉碱:赖氨酸(C:Lys,摩尔比 1:2)、吐温 20 和 γ-CD 与其他同类共溶剂相比,具有更高的(R)-BPFL 产率。此外,基于吐温 20 和 C:Lys(1:2)在提高 BPFO 溶解度和改善细胞通透性方面的优异性能,我们建立了一个包含吐温 20/C:Lys(1:2)的集成反应体系,用于高效生物合成(R)-BPFL。在该协同反应体系中优化了涉及 BPFO 生物还原的关键因素后,BPFO 的负载量提高到 45mM,9h 内的产率达到 90.0%,而在纯水性缓冲液中仅获得 37.6%的产率。这是首次报道利用K. radicincitans 细胞作为新型生物催化剂应用于(R)-BPFL 的制备,开发的包含吐温 20/C:Lys 的协同反应体系在各种手性醇的合成方面具有很大的应用潜力。