SiO2 nanoparticles prepared by the alkoxide hydrolysis precipitation method were covalently immobilized with carbonyl reductase (CR) and formate dehydrogenase (FDH). The nanopartiles immobilized CR and FDH were coupled and used for continuous catalytic conversion of β-hydroxyacetophenone to optical purity (R)-phenyl-ethanediol, and in situ NADH regeneration and recycling analyzed. The results showed that the enzyme loading of nanopartiles immobilized CR and FDH was 3.32 and 5.55 mg/g, their catalytic capability 50%~60% of native enzyme, their optimal reaction pH value 6.5 and 8.5, and their optimal reaction temperature 40 and 45℃, respectively. The production of (R)-phenyl-ethanediol was accumulated as 35.6 g/L, and the productivity of unit nanoparticles 178 g/g, while the coupling system was carried out for 12 batches. The nanopartiles immobilized CR and FDH could be reused after collection with centrifugal separation.