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In this study whey protein concentrate (WPC) at levels (0.1and 4%), soy protein isolate (SPI) at levels (0.1and 2%) and microbial transglutaminase (MTG) at levels (0.1and 2%) were used in low fat ice cream formulation (4% fat) and their effects were evaluated on thephysicochemical and overall acceptance and their optimum amount was determined using response surface methodology. The amount of total solids in all samples was the same (34%). According to the results formulations containing WPC had higher viscosity and acidity but lower overrun, melting resistant and overall acceptance compared to the control sample (<em>P</em><0.01). SPI resulted in greater viscosity and melting resistance but lower overrun and overall acceptance (<em>P</em><0.01). MTG resulted in greater melting resistance, overrun and viscosity of the ice cream samples in comparison to control sample (<em>P</em><0.01). But had no significant effects on overall acceptance and acidity (<em>P</em>>0.05). The result of optimization showed that the best levels of SPI, WPC and MTG were 0.06, 1.37 and 1.19 % respectively. Also in optimum levels, the amount of viscosity, melting resistance, overrun, acidity and overall acceptance were obtained 1700 cp, 52%, 49/54%, 28/5 dornik and 4.25, respectively.