Use of deep eutectic solvents (DESs) to improve biocatalytic asymmetric reduction of 3-chloropropiophenone to (S)-3-chloro-1-phenylpropanol catalyzed by whole-cell of Acetobacter sp. CCTCC M209061 was successfully performed. The cells immobilized on PVA-sodium sulfate exhibited markedly enhanced stability. Diverse DESs as co-solvents manifested significantly different influences on the reaction. Among them, the DES choline chloride/urea ([ChCl][U]) showed the best biocompatibility and moderately increased the cell member permeability, as demonstrated by MAR and flow cytometry assays, and consequently gave the best results. For the bioreduction conducted in the [ChCl][U]-containing system, the optimum [ChCl][U] content, substrate concentration, glucose concentration, pH and temperature were 5% (v/v), 10.0 mmol/L, 60 mmol/L, 5.5 and 30 °C, respectively. Under the optimized conditions, the obtained yield and product e.e. were 82.3% and above 99.0% at a reaction time of 6 h, respectively, and the productivity was 1.37 mmol/L/h. The efficient whole-cell biocatalytic process proved to be feasible on a 500-mL preparative scale. Moreover, the combination of water-immiscible ionic liquid C 4 MIM·PF 6 with [ChCl][U] in a biphasic system further enhanced substrate concentration (16.0 mmol/L), product yield (93.3%) and productivity (1.87 mmol/L/h) significantly, showing to be very promising for biocatalytic synthesis of (S)-3-chloro-1-phenylpropanol with immobilized Acetobacter sp. CCTCC M209061 cells.Acetobacter sp. CCTCC M209061, isolated from Chinese kefir grains by our group, was used as an efficient biocatalyst for highly enantioselective anti-Prelog reduction of prochiral ketones. 11Over the last several years, deep eutectic solvents (DESs) have been considered as another promising alternative to conventional ionic liquids (ILs) as green solvents. 12, 13 DESs are eutectic mixtures composed of two or three cheap and safe components which could associate with each other through hydrogen bond interactions. 14 These solvents not only share the similar characteristics with ILs such as low vapor pressure, non-flammability, thermal stability, but also have some special virtues including relatively low cost, good sustainability, biocompatibility and biodegradability. 15,16 Most importantly, the formation of these solvents is very simple and needs no further purification steps. Up to now, there appears to be some researches related to the applications of DESs as co-solvents in biocatalytic reactions, especially in enzyme-catalyzed reactions. [17][18][19] However, few accounts were published about whole-cell biocatalysis in a In the present study, we attempted for the first time to use various DESs as co-solvents in aqueous system to improve the biocatalytic asymmetric reduction of CPE with immobilized Acetobacter sp. CCTCC M209061 cells (Scheme 1). The biocompatibility of these DESs with the cells and their effects on the cell member integrity were investigated systematically as well as several influential factors on the react...