In this study, the response surface methodology (RSM) was used to optimize the pudding formulation ingredients including the fish/bovine gelatin ratio and cinnamon and clove powder and determine the color and physicochemical and sensory attributes’ change in final pudding product. Experiments were carried out based on a central composite design (CCD). The results showed that by increasing the ratio of fish gelatin to bovine gelatin (FG/BG) up to 3%, the moisture content increased slightly and then decreased significantly. Increasing the cinnamon powder to 0.5% reduced the moisture content. Increasing the FG/BG in the formulation of pudding samples reduced the protein content. The effects of cinnamon and clove powder on the protein content were increasing and decreasing, respectively. By increasing the FG/BG ratio, the samples syneresis showed a significant decrease, while the effects of cinnamon and clove powder on the syneresis were nonsignificant. As the level of cinnamon and clove powder increased, the L* value decreased. Cinnamon and clove powder had a linear effect, and the interaction of gelatins and clove powder had a significant effect on changes in redness. The effects of cinnamon and clove powder on b* value were significant. In terms of sensory evaluation, increasing the cinnamon powder concentration increased the appearance scores, while in the case of fish gelatin, this trend was downward. The linear effect of cinnamon powder on taste was significant, while other variables had no significant effect on the taste of the samples. The sample texture was significantly affected by fish gelatin and clove powder. Increasing FG/BG from 0% to 2.5% increased the texture score, but after this range, a decrease in the texture score was observed. The overall acceptance of samples was more affected by spice powder compared to gelatin. By increasing the cinnamon powder and FG/BG, the overall acceptance increased and decreased, respectively. In conclusion, the optimal FG/BG and cinnamon and clove powder were introduced 1.479%, 0.288%, and 0.619% respectively.