The present study was conducted to evaluate the effect of fish meal replacement with single cell protein on growth performance and proteolytic enzymes activity of pepsin, trypsin and alkaline protease in rainbow trout (Oncorhynchus mykiss) fry (weight: 2.51±0.55g) for 6 weeks. Five experimental diets were prepared with replacement levels of 0 (A), with 25% (B), 50% (C), 75% (D) and 100% (E) of fish meal with microbial protein as isonitrogenous and isoenergetic. Parameters of body weight gain, relative growth rate, protein, lipid and feed efficiency ratio in diet C were significantly the higher than other treatments (p<0.05), while feed conversion ratio was shown the highest and the lowest values in diet C and E, respectively , with a significant difference than those from other treatments (p<0.05). The survival rate was 100 % in the diets. The highest activity of trypsin and alkaline protease enzymes from pyloric caeca and intestine was observed in diet C, which showed a significant difference compared to D and E diets (p<0.05). Also, pepsin had significantly the highest activity in C diet than that from other diets exception for B diet (p<0.05). The findings of growth performance and pepsin, trypsin and alkaline protease activity were revealed the diet containing 50% single cell protein could be appropriate for rainbow trout growth.