Chum salmon cystatin was overexpressed on Saccharomyces cerevisiae YPH 499. At first, the culture condition for the production of recombinant chum salmon cystatin (RC) by S. cerevisiae YPH 499 was optimized in a shake flask using response surface methodology. Three independent variables; medium pH, inducing time, and the amount of inducing assistant, were analyzed to get the optimal condition for the production of RC. The results were fitted to a second-order polynomial equation, in which the determination coefficient ($R^2$) was 0.904. The highest RC production in a shake flask, 0.57 U/mL was obtained at 5.7 of medium pH, 6.7 h of inducing time, and 5.6 g/L of inducing assistant. Based on the results of shake flask, the effects of agitation and aeration rates on the production of RC by S. cerevisiae YPH 499 were determined for scale-up in a fermentor. The highest production of RC in a fermentor, 0.56 U/mL was obtained at 350 rpm of agitation rate and 1.0 vvm of aeration rate. RC at 100 ${mu}g/g$ showed the highest inhibitory activity against the autolysis of Alaska pollock surimi based on the analysis of TCA-soluble peptides.