Optically pure 3-chloro-1-phenyl-1-propanol is an important intermediate for the synthesis of drugs for treating depression. In the present study, biocatalytic asymmetric reduction of 3-chloropropiophenone to (S)-3-chloro-1-phenyl-1-propanol with immobilized Acetobacter sp. CCTCC M209061 cells was successfully conducted in a monophasic aqueous system with high efficiency and selectivity. The immobilized Acetobacter sp. CCTCC M209061 cells exhibited higher stabilities (thermal, pH, and operational stability) than the free cells. In addition, the immobilized microbial cells exhibited a relatively good recyclability; the relative activity of immobilized cells remained over 80%, while that of the free cells under the same condition was less than 20%. In the studied system, glucose was the best co-substrate at an optimum concentration of 50 mmol/L. The optimum buffer pH, reaction temperature, and substrate concentration in this reaction system were 5.5, 30°C, and 3.0 mmol/L, respectively. Under the optimized reaction conditions, the initial reaction rate, yield, and product e.e. were 1.77 mM/h, 88.9%, and above 99.0%, respectively.