The meat industry uses phosphates to improve the water-holding capacity (WHC) of meat products, although excess phosphates can be harmful to human health. In this sense, protein hydrolysates offer an alternative with scientific evidence of improved WHCs. Salmon frames, a byproduct rich in protein, must be processed for recovery. Enzymatic technology allows these proteins to be extracted from muscle, and the sequential batch strategy significantly increases protein nitrogen extraction. This study focused on evaluating the WHC of protein hydrolysates from salmon frames obtained through double- and triple-sequential batches compared to conventional hydrolysis. Hydrolysis was carried out for 3 h at 55 °C with 13 mAU of subtilisin per gram of salmon frames. The WHC of each hydrolysate was measured as the cooking loss using concentrations that varied from 0 to 5% (w/w) in the meat matrix. Compared with those obtained through conventional hydrolysis, the hydrolysates obtained through the strategy of double- and triple-sequence batches demonstrated a 55% and 51% reduction in cooking loss, respectively, when they were applied from 1% by weight in the meat matrix. It is essential to highlight that all hydrolysates had a significantly lower cooking loss (p ≤ 0.05) than that of the positive control (sodium tripolyphosphate [STPP]) at its maximum allowable limit when applied at a concentration of 5% in the meat matrix. These results suggest that the sequential batch strategy represents a promising alternative for further improving the WHC of hydrolysates compared to conventional hydrolysis. It may serve as a viable substitute for polyphosphates.

An experimental methodology to compare the hydrolysis performance of different proteases based on equivalent doses was proposed and validated. The specific activities were quantified via nonlinear fitting of the logarithmic equation to the reaction progress plot. The obtained kinetic constants a were plotted against the protease dose to calculate the specific activities from the slope, resulting in 4214, 1127, 10,277 and 3139 U/g for Alcalase, Flavourzyme, Neutrase and Protamex, respectively. The hydrolysis performances were evaluated using equivalent doses of proteases during the hydrolysis of 50 %(w/w) salmon frame at 50 °C and pH 7.5. Higher degrees of hydrolysis were obtained with Flavourzyme (177 mM and 12 %), whereas greater protein recovery was obtained with Alcalase (55 %). Alcalase was the most cost-efficient protease with 4.3 kg of extracted protein per USD. The presented strategy is an affordable methodology that can be applied to different byproduct-protease systems.

The bioconversion of fish by-products has been evidenced as a sustainable process to convert food waste into high-value products. In the present study, protein hydrolysates were produced from fish by-products by different bioprocesses and evaluated as fertilizers in wheat ( Triticum aestivum L. ) on a nitrogen-equivalent basis. Fish by-products were processed through grinding prior to bioconversion. Enzymatic hydrolysis was performed using Alcalase at 55 °C, pH 6.5, and a 3-h reaction, while microbial conversion was assessed using a lactic culture at 40 °C, pH 6.5, and a 10-day culture. Hydrolysates obtained by enzymatic and microbial bioconversion were evaluated as fertilizers by adding 30 mg after 7 and 14 days to wheat seeds sown under controlled conditions. Protease and microbial hydrolysis generated high concentrations of α -amino groups, yielding 100 mM and 170 mM, respectively. The combined process exhibited a synergistic effect, yielding 226 mM of α -amino groups and 33% of protein recovery. Plant growth assays were conducted under controlled conditions using nitrogen-equivalent doses of each hydrolysate. Microbial and combined enzymatic-microbial hydrolysates generated average plant lengths of 52 cm and 54 cm compared to 44 cm in the control, while plant biomass reached 1.7 g and 2.3 g with microbial and combined enzymatic-microbial hydrolysates compared to 0.7 g in the control. Photosynthetic parameters remained within normal physiological ranges from 2.5 to 3.3 for performance index (PI) and from 0.78 to 0.80 for maximum quantum efficiency (Fv/Fm). The integration of enzymatic and microbial catalysis produced the most effective biostimulant activity, highlighting the value of combining enzymatic specificity with microbial metabolic versatility. These findings support fish-derived protein hydrolysates as efficient and eco-friendly fertilizers that are capable of improving plant growth while contributing to sustainable and integral utilization of natural resources.