Abstract: In this study, the effects of frozenthaw cycles (frozen at -18 ℃ with cycling to 0 ℃ for 12 h and then back to -18 ℃ per 10 days) on molecular structure, free amino groups content, free sulfhydryl groups and farinograph properties of gluten were studied by smallangle Xray scattering (SAXS) and Microdough LAB mixer. The results showed that during the frozenthaw cycles the I(0) of gluten proteins decreased with the increase in freezethaw cycles, especially in the first 60 days storage, the value of I(0) (the scattered intensity implies at zero angle) decreased from 2.75105 for control to 1.75105 for the gluten stored for 60 days, which implies relative molecular mass dropping. The free amino group content changed only slightly, and the free sulphydryl group content increased, indicating that the depolymerisation of the gluten during freezethaw cycling was due to the breakage of disulphide bonds. The farinograph properties of the dough added gluten which after the storage was higher than the one without gluten. The farinograph properties of dough deteriorated with added the frozen stored gluten, especially in the stability time and development time, which related to the relative molecular mass of the additive of the gluten. This suggests that the relative molecular mass and the molecular chain were the decisive factors influencing the quality of the dough for frozen storage.