Towards Zero-Waste Valorization of African Catfish By-Products Through Integrated Biotechnological Processing and Life Cycle Assessment

African catfish (<i>Clarias gariepinus</i>, AC) is one of the most widely farmed freshwater fish species in Central Europe. Processing operations generate up to 55% by-products (BPs), predominantly carcasses rich in proteins, lipids, and minerals. This study develops a comprehensive valorization process for ACBPs to recover gelatin, protein hydrolysate, fish oil, and pigments. The processing protocol consisted of sequential washing, oil extraction, demineralization, and biotechnological treatment to disrupt the collagen quaternary structure. A two-factor experimental design was employed to optimize the processing conditions. The factors included the extraction temperatures of the first (35–45 °C) and second fraction (50–60 °C). We hypothesized that enzymatic conditioning, combined with sequential hot-water extraction, would yield gelatin with properties comparable to those of mammalian- and fish-derived gelatins, while enabling a near-zero-waste process. The integrated process yielded 18.2 ± 1.2% fish oil, 9.8 ± 2.1% protein hydrolysate, 1.7 ± 0.7% pigment extract, and 25.3–37.8% gelatin. Optimal conditions (35 °C/60 °C) produced gelatin with gel strength of 168.8 ± 3.6 Bloom, dynamic viscosity of 2.48 ± 0.02 mPa·s, and yield of 34.76 ± 1.95%. Life cycle assessment (LCA) identified two primary environmental hotspots: water consumption and energy demand. This near-zero-waste biorefinery demonstrates the potential for comprehensive valorization of aquaculture BPs into multiple value-added bioproducts.