Enzymatic hydrolysis of chicken feet for bioactive peptides: method validation and protein quantification
Abstract
Background: Stunting remains a significant public health challenge in Indonesia, affecting 21.5% of children under five years of age, or approximately 6.5 million cases in 2023 and primarily caused by chronic nutritional deficiencies during the critical window of the first 1,000 days of life. Chicken is the second most consumed protein in Indonesia, and produced a lot of by-products with low economic value such as chicken feet. Thus, developing chicken feet bioactive peptide via enzymatic hydrolysis as a treatment for stunting could prove beneficial.
Objective: This study aimed to evaluate the potential of two enzymatic hydrolysis methods—bromelain and a combination of papain and alcalase—for the production of bioactive peptides from chicken feet, and to validate the analytical methods used for protein quantification.
Methods: Chicken feet were hydrolyzed using bromelain, and a papain-alcalase combination. Soluble proteins were quantified using the Biuret assay, and the analytical methods were validated for accuracy, precision, and linearity, with validation criteria including %recovery, relative standard deviation (RSD), and coefficient of determination (R²).
Results: Significant increases in soluble protein content were observed across all treatments (P-value = 0.000). The bromelain method produced higher soluble protein levels, while the papain-alcalase method showed lower protein concentrations, likely due to further degradation of peptides into free amino acids. The analytical methods demonstrated satisfactory performance, with accuracy values within the 98-102% range, RSD values < 2%, and R² values of 0.999, confirming their reliability for protein analysis.
Conclusion: This study demonstrates the potential of enzymatic hydrolysis as a method to make chicken feet, a poultry by-product, into functional ingredients. Further studies in regards of peptide profiles, bioactivity, and functional properties of the hydrolysates is advised for further understanding.
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