In this study, sulfated glycosaminoglycan (GAG) extraction was performed from chicken sternum by enzymatic hydrolysis method and optimum enzymatic hydrolysis process conditions were determined. Response Surface Method was used for the optimization of the process conditions. In order to determine the effects of the selected four parameters as enzyme amount, pH value, extraction temperature and time on GAG yield, a trial design created with Central Composite Design was applied. It is aimed to reach the highest GAG efficiency in the shortest time and by using the least amount of enzyme. The most suitable process conditions for the extraction process were found to be 0.6% enzyme amount, 6.99 pH value, 62.96°C extraction temperature and 10.79 hours extraction time. Under these conditions, the extraction yield value was predicted to be 26.07% and it was confirmed experimentally.