The effect of pretreated method to remove the non-collagenous protein by using alkaline and enzyme Alcalase, as well as the temperature and time for extracting on the properties of gelatin from tra catfish skin were investigated. Yields of gelatin extracted at 70 °C for 1h from pretreated skin by enzyme method (16.2%) was significantly higher than that of the sample by alkaline method (12.14%). However, the gel strength of gelatin from skin treated via enzyme Alcalase was lower than gelatin sample pretreated by alkaline while the turbidity values was higher than gelatin from skin pretreated via alkaline. From SDS-PAGE profile, gelatin from skin pretreated by alkaline consisted of two different α- chains in protein pattern while enzymatic gelatin had low molecular weight peptides. The FT-IR spectra showed the lower wavenumber in amide I and III of enzymatic gelatin in compare to alkaline gelatin by the loss of triple helical structure during enzyme treatment. From the results, the using enzyme for pretreated material has potential to replace the alkaline method for gelatin production with purpose to reduce chemical waste caused serious ecological issues.