Aim An industrial enzyme β-glucanase was used to transfortn notoginsenoside Fe for the first time. Methods Notoginsenoside Fe was isolated from the leave saponin of Panax notoginseng (Burk.) Chen FH. The enzymatically transformed compounds were detected by HPLC and two transformed compounds were identified as 20 (S) -protopanaxadiol-20- O- α-L-arabinofuranosyl ( 1→6 ) - β-gluco- pyranoside, ginsenoside-Mc) and 20(S)-protopanaxadiol-20-O-β-D-glucopyranoside compound-K (C-K) respectively on the basis of their ^1H NMR and ^13 C NMR spectral data. Results Based on the enzymolytic kinetic curve, the transformation rate of notoginsenoside Fe reached 95% after 24 h. Conclusion The enzymatic transformation pathway of notoginsenoside Fe by β-glucanase has been proposed as notoginsenoside Fe→ginsenoside Mc→C-K.