In this study, emulsifiers (sugar ester and sodium stearoyl lactylate), hydrocolloids (guar gum and carboxy methyl cellulose), oxidizers (ascorbic acid and azobisformamide), and enzymes (xylanase and α-amylase) were added into refrigerated dough and revealed their totally different characteristics over 5 days of refrigeration. The optimal refrigerated dough formulation appeared to be the one containing 1% sugar ester, 1% sodium stearoyl lactylate (SSL), 10 mg/kg xylanse and 0.04% ascorbic acid. This optimal formulation predicted a continuous and stable gluten matrix. The secondary structure of refrigerated dough containing optimal formulation remained unchanged while a decrease of 7.00% in α-helix content was observed in control refrigerated dough. It might be ascribed to the enhanced physical intra/intermolecular associations (cross-links or entanglements) of the polymeric chains. These results suggested that the combination of certain additives have a synergistic effect on maintaining the structural characteristics of refrigerated dough.