Microbial whole-cell catalysis has been a popular method for chiral alcohols preparation. In this study, Cyberlindnera saturnus ZJPH1807 was successfully employed as a biocatalyst to reduce 2-chloro-1-(2,4-dichlorophenyl)ethanone (CPE) to (R)-2-chloro-1-(2,4-dichlorophenyl)ethanol ((R)-CPEO), a key chiral intermediate of antifungal drug miconazole. In order to further improve the bioprocess yield, natural deep eutectic solvent and cyclodextrin were introduced into the reaction system as additives. It was found that carnitine/trehalose (C/Tre, 1:1 mol/mol) could increase the permeability of cell membrane appropriately and improve the mass transfer rate. And the addition of methyl-β-cyclodextrin (MCD) could improve the solubility of CPE in buffer solution, simultaneously promoting cofactor recycling. Furthermore, MCD and C/Tre exerted a synergistic effect on the acceleration of CPE bioreduction, producing 91.0% yield in 1.5 h at 35.8 mM CPE in a C/Tre-MCD-containing system, with a 12.4-fold increase in productivity compared to neat buffer solution only. When the substrate loading was increased to 67.1 mM, the yield could also achieve 84.2%. The established bioreduction process was also feasible at 100 mL preparation scale with a 88.5% (R)-CPEO yield. The integration of C/Tre and MCD can significantly improve catalytic efficiency for hydrophobic substrates and be applied in biocatalysis.

微生物全细胞催化一直是制备手性醇的常用方法。在这项研究中， Cyber​​lindnera saturnus ZJPH1807 被成功用作生物催化剂，将 2-chloro-1-(2,4-dichlorophenyl)ethanone (CPE) 还原为 ( R )-2-chloro-1-(2,4-dichlorophenyl)乙醇（（R)-CPEO)，抗真菌药物咪康唑的关键手性中间体。为了进一步提高生物工艺的收率，在反应体系中加入天然低共熔溶剂和环糊精作为添加剂。发现肉碱/海藻糖(C/Tre, 1:1 mol/mol)可以适当增加细胞膜的通透性，提高传质速率。并且甲基-β-环糊精(MCD)的加入可以提高CPE在缓冲溶液中的溶解度，同时促进辅因子的循环利用。此外，MCD 和 C/Tre 对加速 CPE 生物还原发挥协同作用，在含 C/Tre-MCD 的系统中，在 35.8 mM CPE 下 1.5 小时内产生 91.0% 的产率，与产率相比提高了 12.4 倍纯缓冲溶液。当底物负载增加到 67.1 mM 时，收率也可以达到84.2%。所建立的生物还原工艺在 100 mL 制备规模上也是可行的，具有 88.5% (R )-CPEO 产率。C/Tre和MCD的整合可以显着提高疏水底物的催化效率，并应用于生物催化。