By-products of laying hens represent a promising raw material source with a high collagen content, which is currently not adequately used. The aim of the paper is to prepare gelatins from laying hen paws. The purified collagen raw material was processed by a biotechnological process using the food endoprotease Protamex®. After cleavage of the cross-links in the collagen structure, the gelatin was extracted by a batch process with a stirrer in two extraction steps. The influence of the extraction process on the yield of gelatins and on selected qualitative parameters of gelatins was monitored by two-level factor experiments with three selected process factors. The studied factors were: enzyme dosage (0.2−0.8%), enzyme processing time (24−72 h) and gelatin extraction time (30−120 min). After the first extraction step at 75 °C, gelatin was extracted with a yield of 8.2−21.4% and a gel strength of 275−380 Bloom. In the second extraction step at 80−100 °C, it is possible to obtain another portion (3.3−7.7%) of gelatin with a gel strength of 185−273 Bloom. Total extraction efficiency of gelatins prepared from laying hen collagen is almost 30%. The prepared gelatins are of high quality and, under proper extraction conditions, gelatins with a gel strength above 300 Bloom can be prepared, thus equaling commercial beef and pork gelatins of the highest quality. Biotechnological processing of laying hen collagen into gelatins is environmentally friendly.