Extraction and characterization of chicken pepsin with evaluation of its milk-clotting activity in skim milk
https://doi.org/10.21323/2618-9771-2026-9-2-254-260
Abstract
The development of poultry farming creates opportunities to obtain pepsin, a proteolytic enzyme with milk-clotting activity, from the chicken proventriculus (Gallus gallus). This organ, considered a by-product, can serve as a source of proteases, including milk-clotting enzymes that may be used as substitutes for conventional milk-clotting agents in the dairy industry. This study aims to extract pepsin from chicken proventriculus, characterize it, and evaluate its potential use as a rennet substitute. It also attempts to determine the optimal milk clotting conditions (enzyme volume, pH, temperature, and CaCl2 concentration) based on milk flocculation and clotting times. According to the obtained results, the clarified extract of chicken pepsin showed a milk-clotting activity of 63.48 ± 0.21 RU/ml, a specific milk-clotting activity of 1.56 ± 0.19 (RU mg–1), a clotting strength of 12957 ± 399 (SU), a protein content of 43.49 ± 0.75 mg/mL, and a proteolytic activity of 0.91 ± 0.140 μg/mL·h. The highest milk-clotting activity of chicken pepsin was observed between 38 °C and 40 °C, at pH 6.6 and 6.8, with an enzyme volume of 3 mL, and at a CaCl2 concentration of 5 mM. The obtained results indicate the potential use of chicken pepsin as a substitute for rennet in milk clotting. It could also be used in other biological or industrial applications.
About the Authors
D. Bouhadi
Laboratory of Bioconversion, Microbiology Engineering and Health Safety, University Mustapha Stambouli of Mascara
Algeria
Algeria
Djilali Bouhadi, Professor
Postale 305, Route Mamounia, 29000 Mascara
H. Belkhodja
Laboratory of Bioconversion, Microbiology Engineering and Health Safety, University Mustapha Stambouli of Mascara
Algeria
Algeria
Hamza Belkhodja, Associate Professor
Postale 305, Route Mamounia, 29000 Mascara
A. Elouissi
Biological Systems and Geomatics Research Laboratory (LRSBG), University Mustapha Stambouli of Mascara
Algeria
Algeria
Abdelkader Elouissi, Professor
Postale 305, Route Mamounia, 29000, Mascara
Z. Benattouche
Laboratory of Bioconversion, Microbiology Engineering and Health Safety, University Mustapha Stambouli of Mascara
Algeria
Algeria
Zouaoui Benattouche, Professor
Postale 305, Route Mamounia, 29000 Mascara
A. Remil
Laboratory of Organic, Macromolecular Chemistry and Materials, University of Mustapha Stambouli
Algeria
Algeria
Abderrahmane Remil, Associate Professor
Postale 305, Route Mamounia, 29000 Mascara
A. Hariri
Laboratory of Bioconversion, Microbiology Engineering and Health Safety, University Mustapha Stambouli of Mascara
Algeria
Algeria
Ahmed Hariri, Professor
Postale 305, Route Mamounia, 29000 Mascara
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Review
For citations:
Bouhadi D., Belkhodja H., Elouissi A., Benattouche Z., Remil A., Hariri A. Extraction and characterization of chicken pepsin with evaluation of its milk-clotting activity in skim milk. Food systems. 2026;9(2):254-260. https://doi.org/10.21323/2618-9771-2026-9-2-254-260
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