In this study, an NF membrane named S-372 (200 MWCO and the negative charge) was successfully applied in 5'-GMP production. With NF process, 5'-GMP was concentrated from 20.9 g.L-1 to 130.6 g.L-1 and NaCl was desalted from 19.7 g.L-1 to 1.00 g.L-1. The pure water flux could be completely recovered by the alkaline detergent solution with pH 9.0. These results proved that NF process with S-372 could replace vacuum concentration and ethanol crystallization in the conventional process, having advantages of low energy consumption and being environment friendly.