In the present study， the enzymatic hydrolysis resistance of gelatin， modified by transglutaminase， was investigated by single factor test and uniform design. Results showed that the enzymatic hydrolysis resistance of modified gelatin was affected by transglutaminase concentration， treatment pH value， treatment temperature， and treatment time. The optimal treatment conditions for the enzymatic hydrolysis resistance of modified gelatin were as follows： transglutaminase concentration 0.20 g/L， treatment pH 6.0， treatment temperature 44 °C， and treatment time 35 min. The enzymatic hydrolysis resistance of modified gelatin increased 14.44% compared to unmodified gelatin. This study confirmed that the enzymatic hydrolysis resistance of gelatin can be enhanced by transglutaminase modified.