The invention relates to an extraction method for bioactive collagen. The method comprises: a pretreatment step, i.e. pretreating fresh bovine tendon into fragments for standby use; an acid process crude extraction step, i.e. taking 1 part by weight of fragment, adding 4-8 parts by weight of an acetic acid or citric acid solution and conducting stirring, then carrying out coarse filtration, performing high speed centrifugation on a coarse filtrate, then taking the supernatant for standby use; a step of end group removal by an enzymatic process, i.e. adjusting the pH of the supernatant to 9.0, then adding 0.1-0.3% part by weight of Ficus proteinase to conduct enzymolysis digestion; a gradient salting-out step, i.e. using a precipitating agent to conduct multi-gradient salting-out treatment on the solution subjected to enzymolysis to obtain a collagen dissolved solution; and a dialysis step, i.e. loading the collagen dissolved solution into a dialysis bag, using an acetic acid solution and pure water as outer dialysate in order to conduct dialysis, thus obtaining a collagen stock solution with bioactivity. The method provided by the invention combines two existing collagen extraction processes, i.e. the acid process and the enzymatic process, overcomes the low yield defect of the acid process and the incomplete hydrolysis defect of the enzymatic process, and adopts non-animal derived protease, thus reducing the risk of animal origin.