In the present study, to establish the optimum gelatin extraction conditions from pangasius catfish (Pangasius sutchi) bone, Response Surface Methodology (RSM) with a 4-factor, 5-level Central Composite Design (CCD) was conducted. The model equation was proposed with regard to the effects of HCl concentration (%, X 1 ), treatment time (h, X 2 ), extraction temperature (°C, X 3 ) and extraction time (h, X 4 ) as independent variables on the hydroxyproline recovery (%, Y) as dependent variable. X 1 02.74 %, X 2 021.15 h, X 3 074.73°C and X 4 05.26 h were found to be the optimum conditions to obtain the highest hydroxyproline recovery (68.75 %). The properties of optimized catfish bone gelatin were characterized by amino acid analysis, SDS-PAGE, gel strength, TPA and viscosity in comparison to bovine skin gelatin. The result of SDS-PAGE revealed that pangasius catfish bone gelatin consisted of at least 2 different polypeptides (α 1 and α 2 chains) and their cross-linked chains. Moreover, the pangasius catfish bone gelatin was found to contain 17.37 (g/100 g) imino acids (proline and hydroxyproline). Pangasius catfish bone gelatin also indicated physical properties comparable with that of bovine and higher than those from cold water fish gelatin. Based on the results of the present study, there is a potential for exploitation of pangasius catfish bone for gelatin production. Furthermore, RSM provided the best method for optimizing the gelatin extraction parameters.