Exploring Chicken Sternal Cartilage as an Alternative Source of Bioactive Protein Hydrolysates

Open Access

Year : 2025 | Volume : 14 | Issue : 03 | Page : 15 28
    By

    Karina Pinheiro Martins,

  • Thaysa Fernandes Moya Moreira,

  • Anielle de Oliveira,

  • Valéria Maria Costa Teixeira,

  • Odinei Hess Gonçalves,

  • Rafael Porto Ineu,

  • Fernanda Vitória Leimann,

Abstract

There is a growing concern on the correct use and dispose of the by-products of the food industry, and chicken slaughterhouse, such as cartilage, combs, and wattles may be strong candidates as a protein source for bioactive peptides, however it is crucial to understand how the proteolysis process may affect the peptide profile and their bioactivity. The objective of this work was to obtain protein hydrolysates from chicken sternal cartilage using Alcalase and Flavourzyme as catalysts and to determine their antioxidant capacity and acetylcholinesterase (AChE) inhibition potential. The cartilage was hydrolyzed with Alcalase (pH 8, 50°C, 1.8 wt%enzyme:substrate, substrate dry basis) and Flavourzyme (pH 7, 50°C; 1.8 wt%enzyme:substrate, substrate dry basis). Alcalase demonstrated a greater capacity for cartilage digestion (97.42% yield), and the resulting cartilage hydrolyzed a 1.8-fold higher capacity for AChE inhibition than Flavourzyme-catalyzed hydrolysate. On the other hand, Flavourzyme produced a hydrolysate with a 1.3-fold higher capacity on TBARS inhibition (74.2%) than Alcalase. The hydrolysates’ amino acid profiles were delineated via HPLC, revealing the presence of 18 amino acids, notably, tryptophan, tyrosine, and histidine exhibited similar concentrations across the hydrolysates. The tryptophan/LNNA (large neutral amino acids) ratio was elevated in the hydrolysate generated by Flavourzyme, potentially correlating with its heightened TBARS reduction potential. Furthermore, the increased total amino acid content observed in the hydrolysate produced with Alcalase may be linked to its efficacy in AChE inhibition

Keywords: Hydrolysis, Alcalase, Flavourzyme, Tryptophan, large neutral amino acids.

[This article belongs to Research & Reviews : Journal of Food Science & Technology ]

How to cite this article:
Karina Pinheiro Martins, Thaysa Fernandes Moya Moreira, Anielle de Oliveira, Valéria Maria Costa Teixeira, Odinei Hess Gonçalves, Rafael Porto Ineu, Fernanda Vitória Leimann. Exploring Chicken Sternal Cartilage as an Alternative Source of Bioactive Protein Hydrolysates. Research & Reviews : Journal of Food Science & Technology. 2025; 14(03):15-28.
How to cite this URL:
Karina Pinheiro Martins, Thaysa Fernandes Moya Moreira, Anielle de Oliveira, Valéria Maria Costa Teixeira, Odinei Hess Gonçalves, Rafael Porto Ineu, Fernanda Vitória Leimann. Exploring Chicken Sternal Cartilage as an Alternative Source of Bioactive Protein Hydrolysates. Research & Reviews : Journal of Food Science & Technology. 2025; 14(03):15-28. Available from: https://journals.stmjournals.com/rrjofst/article=2025/view=233186


References

1. Romaniw, J.; Inagaki, T. M.; Sá, J. C. de M.; Ramos, F. Bio-Fertilizer Applications from Poultry Slaughterhouses in Subtropical Agriculture – Interactions between Soil Structure and Nitrate Dynamics. Heliyon 2024, 10 (19), e38295. https://doi.org/10.1016/j.heliyon.2024.e38295.
2. Zhao, S.; Thakur, N.; Salama, E.-S.; Zhang, P.; Zhang, L.; Xing, X.; Yue, J.; Song, Z.; Nan, L.; Yujun, S.; Li, X. Potential Applications of Protein-Rich Waste: Progress in Energy Management and Material Recovery. Resour Conserv Recycl 2022, 182, 106315. https://doi.org/10.1016/j.resconrec.2022.106315.
3. Bezerra, T.; Estévez, M.; Lacerda, J. T.; Dias, M.; Juliano, M.; Mendes, M. A.; Morgano, M.; Pacheco, M. T.; Madruga, M. Chicken Combs and Wattles as Sources of Bioactive Peptides: Optimization of Hydrolysis, Identification by LC-ESI-MS2 and Bioactivity Assessment. Molecules 2020, 25 (7), 1698. https://doi.org/10.3390/molecules25071698.
4. Ibarz-Blanch, N.; Alcaide-Hidalgo, J. M.; Cortés-Espinar, A. J.; Albi-Puig, J.; Suárez, M.; Mulero, M.; Morales, D.; Bravo, F. I. Chicken Slaughterhouse By-Products: A Source of Protein Hydrolysates to Manage Non-Communicable Diseases. Trends Food Sci Technol 2023, 139, 104125. https://doi.org/10.1016/j.tifs.2023.104125.
5. Schwartz, S. R.; Hammon, K. A.; Gafner, A.; Dahl, A.; Guttman, N.; Fong, M.; Schauss, A. G. Novel Hydrolyzed Chicken Sternal Cartilage Extract Improves Facial Epidermis and Connective Tissue in Healthy Adult Females: A Randomized, Double-Blind, Placebo-Controlled Trial. Altern Ther Health Med 2019, 25 (5), 12–29.
6. Sakharoff, M. Buteyko Breathing Technique and Ketogenic Diet as Potential Hormetins in Nonpharmacological Metabolic Approaches to Health and Longevity. In The Science of Hormesis in Health and Longevity; Elsevier, 2019; pp 257–274. https://doi.org/10.1016/B978-0-12-814253-0.00023-1.
7. Zhou, D.-D.; Luo, M.; Shang, A.; Mao, Q.-Q.; Li, B.-Y.; Gan, R.-Y.; Li, H.-B. Antioxidant Food Components for the Prevention and Treatment of Cardiovascular Diseases: Effects, Mechanisms, and Clinical Studies. Oxid Med Cell Longev 2021, 2021 (1). https://doi.org/10.1155/2021/6627355.
8. Sonklin, C.; Alashi, M. A.; Laohakunjit, N.; Kerdchoechuen, O.; Aluko, R. E. Identification of Antihypertensive Peptides from Mung Bean Protein Hydrolysate and Their Effects in Spontaneously Hypertensive Rats. J Funct Foods 2020, 64, 103635. https://doi.org/10.1016/j.jff.2019.103635.
9. Asokan, S. M.; Wang, T.; Su, W.-T.; Lin, W.-T. Antidiabetic Effects of a Short Peptide of Potato Protein Hydrolysate in STZ-Induced Diabetic Mice. Nutrients 2019, 11 (4), 779. https://doi.org/10.3390/nu11040779.
10. Daroit, D. J.; Brandelli, A. In Vivo Bioactivities of Food Protein-Derived Peptides – a Current Review. Curr Opin Food Sci 2021, 39, 120–129. https://doi.org/10.1016/j.cofs.2021.01.002.
11. Offengenden, M.; Chakrabarti, S.; Wu, J. Chicken Collagen Hydrolysates Differentially Mediate Anti-Inflammatory Activity and Type I Collagen Synthesis on Human Dermal Fibroblasts. Food Science and Human Wellness 2018, 7 (2), 138–147. https://doi.org/10.1016/j.fshw.2018.02.002.
12. Chen, J.; Yan, Y.; Zhang, L.; Zheng, J.; Guo, J.; Li, R.; Zeng, J. Purification of Novel Antioxidant Peptides from Myofibrillar Protein Hydrolysate of Chicken Breast and Their Antioxidant Potential in Chemical and H 2 O 2 -Stressed Cell Systems. Food Funct 2021, 12 (11), 4897–4908. https://doi.org/10.1039/D1FO00579K.
13. Aloysius, T.; Carvajal, A.; Slizyte, R.; Skorve, J.; Berge, R.; Bjørndal, B. Chicken Protein Hydrolysates Have Anti-Inflammatory Effects on High-Fat Diet Induced Obesity in Mice. Medicines 2018, 6 (1), 5. https://doi.org/10.3390/medicines6010005.
14. Brandao-Rangel, M. A. R.; Oliveira, C. R.; da Silva Olímpio, F. R.; Aimbire, F.; Mateus-Silva, J. R.; Chaluppe, F. A.; Vieira, R. P. Hydrolyzed Collagen Induces an Anti-Inflammatory Response That Induces Proliferation of Skin Fibroblast and Keratinocytes. Nutrients 2022, 14 (23), 4975. https://doi.org/10.3390/nu14234975.
15. Zague, V. A New View Concerning the Effects of Collagen Hydrolysate Intake on Skin Properties. Arch Dermatol Res 2008, 300 (9), 479–483. https://doi.org/10.1007/s00403-008-0888-4.
16. Soniwala, S.; Scinto, K. I.; Schott, E. M.; Stolarczyk, A. E.; Villani, D. A.; Dar, Q.-A.; Grier, A.; Ketz, J. P.; Gill, S. R.; Mooney, R. A.; Prawitt, J.; Zuscik, M. J. Oral Hydrolyzed Type 2 Collagen Protects against the OA of Obesity and Mitigates Obese Gut Microbiome Dysbiosis. Osteoarthritis Cartilage 2018, 26, S173–S174. https://doi.org/10.1016/j.joca.2018.02.377.
17. Terry, A. V.; Buccafusco, J. J. The Cholinergic Hypothesis of Age and Alzheimer’s Disease-Related Cognitive Deficits: Recent Challenges and Their Implications for Novel Drug Development. Journal of Pharmacology and Experimental Therapeutics 2003, 306 (3), 821–827. https://doi.org/10.1124/jpet.102.041616.
18. Kim, D.; Kim, Y. H. B.; Ham, J.-S.; Lee, S. K.; Jang, A. Pig Skin Gelatin Hydrolysates Attenuate Acetylcholine Esterase Activity and Scopolamine-Induced Impairment of Memory and Learning Ability of Mice. Food Sci Anim Resour 2020, 40 (2), 183–196. https://doi.org/10.5851/kosfa.2020.e3.
19. Wu, Y.-H. S.; Lin, Y.-L.; Yang, W.-Y.; Wang, S.-Y.; Chen, Y.-C. Pepsin-Digested Chicken-Liver Hydrolysate Attenuates Hepatosteatosis by Relieving Hepatic and Peripheral Insulin Resistance in Long-Term High-Fat Dietary Habit. J Food Drug Anal 2021, 29 (2), 376–389. https://doi.org/10.38212/2224-6614.3351.
20. Chen, C.-C.; Chang, S.-S.; Chang, C.-H.; Hu, C.-C.; Nakao, Y.; Yong, S. M.; Mandy, Y. L. O.; Lim, C. J.; Shim, E. K.-S.; Shih, H.-N. Randomized, Double-Blind, Four-Arm Pilot Study on the Effects of Chicken Essence and Type II Collagen Hydrolysate on Joint, Bone, and Muscle Functions. Nutr J 2023, 22 (1), 17. https://doi.org/10.1186/s12937-023-00837-w.
21. Cao, H.; Xu, S.-Y. Purification and Characterization of Type II Collagen from Chick Sternal Cartilage. Food Chem 2008, 108 (2), 439–445. https://doi.org/10.1016/j.foodchem.2007.09.022.
22. Saidi, S.; Belleville, M.-P.; Deratani, A.; Amar, R. Ben. Optimization of Peptide Production by Enzymatic Hydrolysis of Tuna Dark Muscle By-Product Using Commercial Proteases. Afr J Biotechnol 2013, 12 (13), 1533–1547. https://doi.org/10.5897/AJB12.2745.
23. Alahmad, K.; Noman, A.; Xia, W.; Jiang, Q.; Xu, Y. Influence of the Enzymatic Hydrolysis Using Flavourzyme Enzyme on Functional, Secondary Structure, and Antioxidant Characteristics of Protein Hydrolysates Produced from Bighead Carp (Hypophthalmichthys Nobilis). Molecules 2023, 28 (2). https://doi.org/10.3390/molecules28020519.
24. Lutz, I. A. Métodos Físico-Químicos Para Análise de Alimentos; 2008.
25. Moreira, T. F. M.; Pessoa, L. G. A.; Seixas, F. A. V.; Ineu, R. P.; Gonçalves, O. H.; Leimann, F. V.; Ribeiro, R. P. Chemometric Evaluation of Enzymatic Hydrolysis in the Production of Fish Protein Hydrolysates with Acetylcholinesterase Inhibitory Activity. Food Chem 2022, 367, 130728. https://doi.org/10.1016/j.foodchem.2021.130728.
26. Hagen, S. R.; Frost, B.; Augustin, J. Precolumn Phenylisothiocyanate Derivatization and Liquid Chromatography of Amino Acids in Food. Journal of the Association of Official Analytical Chemists 1989, 72 (6), 912–916.
27. White, J. A.; Hart, R. J.; Fry, J. C. An Evaluation of the Waters Pico-Tag System for the Amino-Acid Analysis of Food Materials. J Anal Methods Chem 1986, 8, 170–177.
28. White, J. A.; Hart, R. J.; Fry, J. C. An Evaluation of the Waters Pico-Tag System for the Amino-Acid Analysis of Food Materials. Journal of Automatic Chemistry 1986, 8 (4), 170–177.
29. Gatti, R.; Gioia, M. G.; Andreatta, P.; Pentassuglia, G. HPLC-Fluorescence Determination of Amino Acids in Pharmaceuticals after Pre-Column Derivatization with Phanquinone. In Journal of Pharmaceutical and Biomedical Analysis; 2004; Vol. 35, pp 339–348. https://doi.org/10.1016/S0731-7085(03)00584-3.
30. Brand-Williams, W.; Cuvelier, M. E.; Berset, C. Use of a Free Radical Method to Evaluate Antioxidant Activity. LWT – Food Science and Technology 1995, 28 (1), 25–30. https://doi.org/10.1016/S0023-6438(95)80008-5.
31. Thaipong, K.; Boonprakob, U.; Crosby, K.; Cisneros-Zevallos, L.; Hawkins Byrne, D. Comparison of ABTS, DPPH, FRAP, and ORAC Assays for Estimating Antioxidant Activity from Guava Fruit Extracts. Journal of Food Composition and Analysis 2006, 19 (6–7), 669–675. https://doi.org/10.1016/j.jfca.2006.01.003.
32. Ueda, J. M.; Griebler, K. R.; Finimundy, T. C.; Rodrigues, D. B.; Veríssimo, L.; Pires, T. C. S. P.; Gonçalves, J.; Fernandes, I. P.; Pereira, E.; Barros, L.; Heleno, S. A.; Calhelha, R. C. Polyphenol Composition by HPLC-DAD-(ESI-)MS/MS and Bioactivities of Extracts from Grape Agri-Food Wastes. Molecules 2023, 28 (21), 7368. https://doi.org/10.3390/molecules28217368.
33. Ellman, G. L.; Courtney, K. D.; Andres, V.; Featherstone, R. M. A New and Rapid Colorimetric Determination of Acetylcholinesterase Activity. Biochem Pharmacol 1961, 7 (2), 88–95. https://doi.org/10.1016/0006-2952(61)90145-9.
34. Akram, A. N.; Zhang, C. Effect of Ultrasonication on the Yield, Functional and Physicochemical Characteristics of Collagen-II from Chicken Sternal Cartilage. Food Chem 2020, 307, 125544. https://doi.org/10.1016/j.foodchem.2019.125544.
35. Shen, Q.; Zhang, C.; Jia, W.; Qin, X.; Cui, Z.; Mo, H.; Richel, A. Co-Production of Chondroitin Sulfate and Peptide from Liquefied Chicken Sternal Cartilage by Hot-Pressure. Carbohydr Polym 2019, 222, 115015. https://doi.org/10.1016/j.carbpol.2019.115015.
36. Shen, Q.; Zhang, C.; Jia, W.; Qin, X.; Xu, X.; Ye, M.; Mo, H.; Richel, A. Liquefaction of Chicken Sternal Cartilage by Steam Explosion to Isolate Chondroitin Sulfate. Carbohydr Polym 2019, 215, 73–81. https://doi.org/10.1016/j.carbpol.2019.03.032.
37. Ren, J.; Song, C.-L.; Zhang, H.-Y.; Kopparapu, N.-K.; Zheng, X.-Q. Effect of Hydrolysis Degree on Structural and Interfacial Properties of Sunflower Protein Isolates. J Food Process Preserv 2017, 41 (1), e13092. https://doi.org/10.1111/jfpp.13092.
38. Alahmad, K.; Xia, W.; Jiang, Q.; Xu, Y. Effect of the Degree of Hydrolysis on Nutritional, Functional, and Morphological Characteristics of Protein Hydrolysate Produced from Bighead Carp (Hypophthalmichthys Nobilis) Using Ficin Enzyme. Foods 2022, 11 (9), 1320. https://doi.org/10.3390/foods11091320.
39. Ovissipour, M.; Kenari, A. A.; Motamedzadegan, A.; Rasco, B.; Nazari, R. M. Optimization of Protein Recovery During Hydrolysis of Yellowfin Tuna ( Thunnus Albacares ) Visceral Proteins. Journal of Aquatic Food Product Technology 2011, 20 (2), 148–159. https://doi.org/10.1080/10498850.2010.548910.
40. Chalamaiah, M.; Dinesh kumar, B.; Hemalatha, R.; Jyothirmayi, T. Fish Protein Hydrolysates: Proximate Composition, Amino Acid Composition, Antioxidant Activities and Applications: A Review. Food Chem 2012, 135 (4), 3020–3038. https://doi.org/10.1016/j.foodchem.2012.06.100.
41. Xiao, Q.; Woo, M. W.; Hu, J.; Xiong, H.; Zhao, Q. The Role of Heating Time on the Characteristics, Functional Properties and Antioxidant Activity of Enzyme-Hydrolyzed Rice Proteins-Glucose Maillard Reaction Products. Food Biosci 2021, 43, 101225. https://doi.org/10.1016/j.fbio.2021.101225.
42. van der Heijden, F. M. M. A.; Fekkes, D.; Tuinier, S.; Sijben, A. E. S.; Kahn, R. S.; Verhoeven, W. M. A. Amino Acids in Schizophrenia: Evidence for Lower Tryptophan Availability during Treatment with Atypical Antipsychotics? J Neural Transm 2005, 112 (4), 577–585. https://doi.org/10.1007/s00702-004-0200-5.
43. Kikuchi, A. M.; Tanabe, A.; Iwahori, Y. A Systematic Review of the Effect of L-Tryptophan Supplementation on Mood and Emotional Functioning. J Diet Suppl 2021, 18 (3), 316–333. https://doi.org/10.1080/19390211.2020.1746725.
44. Zahar, S.; Schneider, N.; Makwana, A.; Chapman, S.; Corthesy, J.; Amico, M.; Hudry, J. Dietary Tryptophan-Rich Protein Hydrolysate Can Acutely Impact Physiological and Psychological Measures of Mood and Stress in Healthy Adults. Nutr Neurosci 2023, 26 (4), 303–312. https://doi.org/10.1080/1028415X.2022.2047435.
45. Templeman, J. R.; Thornton, E.; Cargo-Froom, C.; Squires, E. J.; Swanson, K. S.; Shoveller, A. K. Effects of Incremental Exercise and Dietary Tryptophan Supplementation on the Amino Acid Metabolism, Serotonin Status, Stool Quality, Fecal Metabolites, and Body Composition of Mid-Distance Training Sled Dogs. J Anim Sci 2020, 98 (5). https://doi.org/10.1093/jas/skaa128.
46. Chakraborty, S.; Singh, O. P.; Dasgupta, A.; Mandal, N.; Das, H. N. Correlation between Lipid Peroxidation-Induced TBARS Level and Disease Severity in Obsessive–Compulsive Disorder. Prog Neuropsychopharmacol Biol Psychiatry 2009, 33 (2), 363–366. https://doi.org/10.1016/j.pnpbp.2009.01.001.
47. Karoud, W.; Brahmi, N.; Hamed, H.; Allagui, M. S.; Bougatef, A.; Sila, A. Hake Heads Proteins Hydrolysates as a Source of Bioactive Peptides with Prevention against Hypercholesterolemia in Mice: The Involvement of Oxidative Dysfunction in Hepatic Tissues. Food Chemistry Advances 2023, 2, 100266. https://doi.org/10.1016/j.focha.2023.100266.
48. de Medeiros, B. Z.; Wessler, L. B.; Duarte, M. B.; Lemos, I. S.; Candiotto, G.; Canarim, R. O.; dos Santos, P. C. L.; Torres, C. A.; Scaini, G.; Rico, E. P.; Generoso, J. S.; Streck, E. L. Exposure to Leucine Induces Oxidative Stress in the Brain of Zebrafish. Metab Brain Dis 2022, 37 (4), 1155–1161. https://doi.org/10.1007/s11011-022-00934-5.
49. Barschak, A. G.; Sitta, A.; Deon, M.; de Oliveira, M. H.; Haeser, A.; Dutra-Filho, C. S.; Wajner, M.; Vargas, C. R. Evidence That Oxidative Stress Is Increased in Plasma from Patients with Maple Syrup Urine Disease. Metab Brain Dis 2006, 21 (4), 279–286. https://doi.org/10.1007/s11011-006-9030-5.
50. Moreira, T. F. M.; Pessoa, L. G. A.; Seixas, F. A. V.; Ineu, R. P.; Gonçalves, O. H.; Leimann, F. V.; Ribeiro, R. P. Chemometric Evaluation of Enzymatic Hydrolysis in the Production of Fish Protein Hydrolysates with Acetylcholinesterase Inhibitory Activity. Food Chem 2022, 367. https://doi.org/10.1016/j.foodchem.2021.130728.
51. Halim, N. R. A.; Azlan, A.; Yusof, H. M.; Sarbon, N. M. Antioxidant and Anticancer Activities of Enzymatic Eel (Monopterus Sp) Protein Hydrolysate as Influenced by Different Molecular Weight. Biocatal Agric Biotechnol 2018, 16, 10–16. https://doi.org/10.1016/j.bcab.2018.06.006.
52. Halim, N. R. A.; Yusof, H. M.; Sarbon, N. M. Functional and Bioactive Properties of Fish Protein Hydolysates and Peptides: A Comprehensive Review. Trends Food Sci Technol 2016, 51, 24–33. https://doi.org/10.1016/j.tifs.2016.02.007.
53. Chi, C. F.; Cao, Z. H.; Wang, B.; Hu, F. Y.; Li, Z. R.; Zhang, B. Antioxidant and Functional Properties of Collagen Hydrolysates from Spanish Mackerel Skin as Influenced by Average Molecular Weight. Molecules 2014, 19 (8), 11211–11230. https://doi.org/10.3390/molecules190811211.
54. Jongsriwattanaporn, M.; Chandarajoti, K.; Petrat, W.; Kara, J.; Plubrukarn, A.; Premchai, P.; Lomlim, L. In Vitro Anti-Oxidation and Anti-Cholinesterase Activities of Tuna and Chicken Hydrolysates and Their Maillard Reaction Products. Journal of Health Science and Medical Research 2023, 2023998. https://doi.org/10.31584/jhsmr.2023998.

Regular Issue Open Access Original Research
Volume 14
Issue 03
Received 12/04/2025
Accepted 06/05/2025
Published 27/11/2025
Publication Time 229 Days
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