The cofactor recycling and reutilization of carbonyl reductase catalysis in Candida parapsilosis (C. parapsilosis) was reinforced by coupling with permeabilized cell of Candida boidinii (C. boidinii). Thus an effective permeabilized cell coupling system was constructed for preparation chiral (R)-phenethanol. The experiment results were as follows: substrate and production infiltration ability of permeabilized C. parapsilosis were enhanced, its substrate conversion rate was improved from 33.5% to 43.8%; the substrate conversion rate was further improved from 43.8 to 52.1% by adding cofactor in addition; the cofactor recycling efficiency could be greatly enhanced by coupling with permeabilized C. boidinii with adding cofactor in addition, the substrate conversion rate could reach as 71.8%. The permeabilized cell coupling system reaction conditions were optimized as: when the C. parapsilosis cell mass was 0.2 g, the cell mass ratio of C. parapsilosis and C. boidinii was 1.5:1, substrate concentration was 10 mmol/L, cofactor concentration was 1% of substrate concentration (as 0.1 mmol/L), its substrate conversion rate could reach as 78.2%. The cells were re-collected by centrifugation for multibatches reaction. The substrate conversion rate could still keep at 55% when the coupling system was reacted for 12 batches, the total substrate conversion rate was 70.3%, the production was determined as (R)-phenethanol, its enantiomeric excess(e.e.) was 98.2%.