This study was conducted to identify the factors affecting the soluble protein extraction from cultured snakehead fish muscle, a protein that has high bioavailability and nutritional value. The study investigated the ratio of fish muscle and water (from 1:1 to 1:5 w/v) as well as suitable salt (NaCl) concentration (from 0.05 M to 0.25 M) for soluble protein extraction to be highly effective. Research has applied a response surface methodology using a central composite design to optimize the effect of temperature and time on soluble protein extraction. In addition, the study investigated the effect of pH of salt solution (pH 2 to pH 12) on the efficiency of soluble protein extraction from cultured snakehead fish muscle. As a result, the use of 0.15 M NaCl with 1:2 ratio of fish muscle and salt solution gave high soluble protein extraction efficiency. The optimum extraction temperature and time showed that, the optimum condition for solubilization of soluble protein was determined as 34.8°C and 15.2 min respectively, and the appropriate pH value was also determined as pH 10. At the optimum extraction conditions, soluble protein recovery calculated by total protein of fish muscle were 14.39±0.18%.