Biotechnological valorization of chicken processing by-products for sustainable peptone production

Abstract

In this study, it was aimed to produce a sustainable chicken peptone through the biotechnological valorization of poultry processing by-products via enzymatic hydrolysis using alcalase. The effective utilization of protein-rich wastes generated by increasing animal production activities, without causing environmental harm, is of great importance for sustainable production approaches and circular economy principles. In this context, converting such wastes into value-added bioprocess products is noteworthy for both reducing environmental burden and providing economic benefits. Response Surface Methodology was employed to optimize the enzymatic hydrolysis process, and the effects of enzyme concentration and hydrolysis time on peptone yield and protein content were systematically investigated. The optimum conditions were determined as 0.5% enzyme concentration and 12 hours of hydrolysis, under which the highest peptone yield (60%) and protein content (92.00%) were achieved. Chemical characterization results demonstrated that the produced chicken peptone has high total nitrogen (16.22%), free amino nitrogen (4.51%), and soluble protein content. Structural analyses revealed that the peptone predominantly consists of low molecular weight peptide fractions, with a high degree of hydrolysis (28.56%) and an average molecular weight of 418 Da, indicating high bioavailability and rapid metabolizability. Furthermore, the amino acid composition was rich in glutamic acid, aspartic acid, alanine, and glycine, which are critical for microbial metabolism. Evaluation of the produced peptone as a microbial growth medium showed comparable, and in some cases superior, growth performance to commercial peptones in cultures of Escherichia coli and Bacillus subtilis. This performance is associated with its balanced amino acid profile, low molecular weight peptide content, and high bioavailability. Overall, the findings demonstrate that poultry processing by-products are a promising alternative resource for the sustainable, economical, and environmentally friendly production of high value-added peptones, with significant potential for industrial-scale applications.

Keywords

• Chicken processing by-products
• hydrolysis
• peptone
• Escherichia coli
• Bacillus subtilis

Tavuk işleme atıklarının sürdürülebilir biyoteknolojik dönüştürülmesiyle pepton üretimi

Abstract

Bu çalışmada, tavuk işleme atıklarının alkalaz enzimi kullanılarak enzimatik hidroliz yoluyla biyoteknolojik olarak değerlendirilmesiyle sürdürülebilir bir tavuk peptonu üretimi amaçlanmıştır. Artan hayvansal üretim faaliyetleri sonucunda ortaya çıkan protein açısından zengin atıkların çevreye zarar vermeden değerlendirilmesi, sürdürülebilir üretim yaklaşımları ve döngüsel ekonomi perspektifi açısından büyük önem taşımaktadır. Bu kapsamda, söz konusu atıkların katma değerli biyoproses ürünlerine dönüştürülmesi hem çevresel yükün azaltılması hem de ekonomik fayda sağlanması açısından dikkat çekmektedir. Enzimatik hidroliz sürecinin optimizasyonu için Yanıt Yüzey Metodolojisi uygulanmış; enzim konsantrasyonu ve hidroliz süresinin pepton verimi ve protein içeriği üzerindeki etkileri sistematik olarak incelenmiştir. Optimum koşullar %0,5 enzim konsantrasyonu ve 12 saat hidroliz süresi olarak belirlenmiş ve bu şartlarda en yüksek pepton verimi (%60) ile protein oranı (%92,00) elde edilmiştir. Kimyasal karakterizasyon sonuçları, üretilen tavuk peptonunun yüksek toplam azot (%16,22), serbest amino azot (%4,51) ve çözünür protein içeriğine sahip olduğunu göstermiştir. Yapısal analizler, peptonun yüksek hidroliz derecesi (%28,56) ve ortalama 418 Da molekül ağırlığı ile ağırlıklı olarak düşük molekül ağırlıklı peptit fraksiyonlarından oluştuğunu ortaya koymuştur. Bu durum, ürünün biyoyararlanım potansiyelinin yüksek olduğuna ve hızlı metabolize edilebilirliğine işaret etmektedir. Ayrıca amino asit kompozisyonunun, mikrobiyal metabolizma açısından kritik öneme sahip glutamik asit, aspartik asit, alanin ve glisin bakımından zengin olduğu belirlenmiştir. Üretilen peptonun mikrobiyal büyüme ortamı olarak etkinliği değerlendirildiğinde, Escherichia coli ve Bacillus subtilis kültürlerinde ticari peptonlara eşdeğer hatta bazı durumlarda daha yüksek büyüme performansı sağladığı saptanmıştır. Bu durum, peptonun dengeli amino asit profili, düşük molekül ağırlıklı peptit içeriği ve yüksek biyoyararlanımı ile doğrudan ilişkilendirilmektedir. Elde edilen bulgular, tavuk işleme atıklarının yüksek katma değerli biyoproses ürünlerine dönüştürülmesinde etkili bir yaklaşım sunduğunu ve sürdürülebilir, ekonomik ve çevre dostu pepton üretimi için önemli bir alternatif kaynak oluşturduğunu ortaya koymaktadır. Ayrıca, geliştirilen bu yaklaşımın endüstriyel ölçekte uygulanabilirliği açısından da önemli bir potansiyel taşıdığı değerlendirilmektedir.

Keywords

• Tavuk işleme atıkları
• hidroliz
• pepton
• Escherichia coli
• Bacillus subtilis

References

1. Beyaz Et Sanayicileri ve Damızlıkçıları Birliği Derneği. (2024). Kanatlı beyaz et sektörü üretim verileri [Poultry white meat sector production data]. BESD-BİR. https://www.besd-bir.org/sektor-verileri/
2. Chen, J., Li, L., Yi, R., Xu, N., Gao, R., & Hong, B. (2016). Extraction and characterization of acid-soluble collagen from scales and skin of tilapia (Oreochromis niloticus), LWT - Food Science and Technology, 66(9), 453-459. https://doi.org/10.1016/j.lwt.2015.10.070
3. Cui, Q., Li, Y., Li, T., Yu, J., Shen, G., Sun, X., Zhou, M., & Zhang, Z. (2024). Characterization of Peptide Profiles and the Hypoallergenic and High Antioxidant Activity of Whey Protein Hydrolysate Prepared Using Different Hydrolysis Modes. Foods, 13(18), 2978. https://doi.org/10.3390/foods13182978
4. Cunniff, P. (Ed.). (1997). Official methods of analysis of AOAC International (16th ed., 4th rev.). AOAC International. https://doi.org/10.1093/jaoac/80.6.127A
5. Davami, F., Eghbalpour, F., Nematollahi, L., Barkhordari, F., & Mahboudi, F. (2015). Effects of peptone supplementation in different culture media on growth, metabolic pathway and productivity of CHO DG44 cells; A new insight into amino acid profiles. Iranian Biomedical Journal, 19(4), 194–205. https://doi.org/10.7508/ibj.2015.04.002
6. Deepachandi, B., Weerasinghe, S., Andrahennadi, T. P., Karunaweera, N.D., Wickramarachchi, N., Soysa, P., & Siriwardana, Y. (2020). Quantification of soluble or insoluble fractions of Leishmania parasite proteins in microvolume applications: A simplification to standard Lowry assay. International Journal of Analytical Chemistry, 2020, Article 6129132. https://doi.org/10.1155/2020/6129132
7. Drabold, E. T., Sakhakarmy, M., Shanmugam, S. R., Adhikari, S., Arthur, W., Rudar, M., Boersma, M., Wang, Q., & Higgins, B. T. (2025). Thermal hydrolysis of poultry byproducts for the production of microbial media. Scientific Reports, 15, Article 6107. https://doi.org/10.1038/s41598-025-90411-7
8. Fallah, M., Bahram, S., & Javadian, S. R. (2015). Fish peptone development using enzymatic hydrolysis of silver carp by-products as a nitrogen source in Staphylococcus aureus media. Food science & nutrition, 3(2), 153–157. https://doi.org/10.1002/fsn3.198

9. Food and Agriculture Organization of the United Nations. (2014). Poultry development review. https://www.fao.org/documents/card/en/c/i3531e
10. Göçer, M., Yanar, Y., & Aydın, M. (2023). Determination of amino acid profile and some characteristics of collagen extracted from skin and bone of Mangar (Luciobarbus esocinus Heckel, 1843). LimnoFish, 9(2), 94–107. https://doi.org/10.17216/LimnoFish.1213720
11. Hubálek Z. (2003). Protectants used in the cryopreservation of microorganisms. Cryobiology, 46(3), 205–229. https://doi.org/10.1016/s0011-2240(03)00046-4
12. Islam, M., Huang, Y., Islam, S., Fan, B., Tong, L., & Wang, F. (2022). Influence of the Degree of Hydrolysis on Functional Properties and Antioxidant Activity of Enzymatic Soybean Protein Hydrolysates. Molecules, 27(18), 6110. https://doi.org/10.3390/molecules27186110
13. Kosasih, W., Ratnaningrum, D., Endah, E. S., Pudjiraharti, S., & Priatni, S. (2018). The use of papain enzyme in fish peptone processing. In IOP Conference Series: Earth and Environmental Science, 160(1), Article 012007. IOP Publishing Ltd. https://doi.org/10.1088/1755-1315/160/1/012007
14. Kuncorojakti, S., Delaiah, D., Aswin, A., Puspitasari, Y., Damayanti, Y., Susilowati, H., Diyantoro, Hamid, I., Arif, M., Suwarno, & Rodprasert, W. (2024). Development of peptone-based serum-free media to support Vero CCL-81 cell proliferation and optimize SARS-CoV2 viral production. Heliyon, 10(13), e41077. https://doi.org/10.1016/j.heliyon.2024.e41077
15. Liu, G., Tiang, M. F., Ma, S., Wei, Z., Liang, X., Sajab, M. S., Abdul, P. M., Zhou, X., Ma, Z., & Ding, G. (2024). An alternative peptone preparation using Hermetia illucens (Black soldier fly) hydrolysis: process optimization and performance evaluation. PeerJ, 12, e16995. https://doi.org/10.7717/peerj.16995
16. López-Morales, C. A., Vázquez-Leyva, S., Vallejo-Castillo, L., Carballo-Uicab, G., Muñoz-García, L., Herbert-Pucheta, J. E., Zepeda-Vallejo, L. G., Velasco-Velázquez, M., Pavón, L., Pérez-Tapia, S. M., & Medina-Rivero, E. (2019). Determination of peptide profile consistency and safety of collagen hydrolysates as quality attributes. Journal of Food Science, 84(3), 430–439. https://doi.org/10.1111/1750-3841.14466
17. Nakamura, A., Takahashi, H., Sulaiman, S., Phraephaisarn, C., Keeratipibul, S., Kuda, T., & Kimura, B. (2021). Evaluation of peptones from chicken waste as a nitrogen source for micro-organisms. Letters in applied microbiology, 72(4), 408–414. https://doi.org/10.1111/lam.13428
18. Nielsen, P. M., Petersen, D., & Dambmann, C. (2001). Improved method for determining food protein degree of hydrolysis. Journal of Food Science, 66(5), 642–646. https://doi.org/10.1111/j.1365-2621.2001.tb04614.x
19. Petrova, I., Tolstorebrov, I., Zhivlyantseva, I., & Eikevik, T. M. (2021). Utilization of fish protein hydrolysates as peptones for microbiological culture medias, Food Bioscience, 42, 101063. https://doi.org/10.1016/j.fbio.2021.101063.
20. Priatni, S., Kosasih, W., Budiwati, T., & Ratnaningrum, D. (2017). Production of peptone from boso fish (Oxyeleotris marmorata) for bacterial growth medium. IOP Conference Series: Earth and Environmental Science, 60(1), Article 012009. https://doi.org/10.1088/1755-1315/60/1/012009
21. Rezaee, N., Hasanvand, P., Bagheri Lotfabad, T., Heydarinasab, A., Khodabandeh, M., & Yaghmaei, S. (2022). Study on the use of bovine blood protein hydrolysate as a peptone in microbial culture media. Preparative Biochemistry & Biotechnology, 53(5), 531–539. https://doi.org/10.1080/10826068.2022.2119577
22. Saeed, H. M., Ragaey, A. M., Samy, Z. A., Awad, H. M., & El-Toukhy, N. M. (2025). Optimization and characterization studies of poultry waste valorization for peptone production using a newly Egyptian Bacillus subtilis strain. AMB Express, 15, Article 9. https://doi.org/10.1186/s13568-024-01794-1
23. Seidavi, A., Zaker, H., & Scanes, C. G. (2019). Present and potential impacts of waste from poultry production on the environment. World's Poultry Science Journal, 75(1), 1–14. https://doi.org/10.1017/S0043933918000922
24. Setijawati, D., Jaziri, A., Yufidasari, H., Wardani, D., Dwi Pratomo, M., Ersyah, D., & Huda, N. (2019). Characteristics of Peptone from the Mackerel, Scomber japonicus Head by-Product as Bacterial Growth Media. Bioscience Biotechnology Research Communications, 12, 829. https://doi.org/10.21786/bbrc/12.4/1
25. Taskin, M., Sisman, T., Erdal, S., & Inan, O. (2011). Use of waste chicken feathers as peptone for production of carotenoids in submerged culture of Rhodotorula glutinis MT-5. European Food Research and Technology, 233(4), 657–665. https://doi.org/10.1007/s00217-011-1561-2
26. Trivedi, M., Branton, A., Trivedi, D., Nayak, G., Mishra, R., & Jana, S. (2015). Physicochemical evaluation of biofield treated peptone and Malmgren modified terrestrial orchid medium. American Journal of Bioscience and Bioengineering, 3(6), 169–177. https://doi.org/10.11648/j.bio.20150306.15
27. Türkiye İstatistik Kurumu. (2024). Kümes hayvancılığı üretimi [Poultry production]. TÜİK Haber Bülteni. https://data.tuik.gov.tr/Bulten/Index?p=Kumes-Hayvanciligi-Uretimi-Mayis-2024-49659
28. U.S. Department of Agriculture, Foreign Agricultural Service. (2025, April 10). Livestock and poultry: World markets and trade. https://www.fas.usda.gov/data/livestock-and-poultry-world-markets-and-trade
29. Vasileva-Tonkova, E., Nustorova, M., & Gushterova, A. (2007). New protein hydrolysates from collagen wastes used as peptone for bacterial growth. Current microbiology, 54(1), 54–57. https://doi.org/10.1007/s00284-006-0308-y
30. Vázquez, J. A., Durán, A. I., Menduíña, A., & Nogueira, M. (2020). Biotechnological Valorization of Food Marine Wastes: Microbial Productions on Peptones Obtained from Aquaculture By-Products. Biomolecules, 10(8), 1184. https://doi.org/10.3390/biom10081184
31. Wubshet, S. G., Måge, I., Böcker, U., Lindberg, D., Knutsen, S. H., Rieder, A., Airado-Rodríguez, D., & Afseth, N. K. (2017). FTIR as a rapid tool for the evaluation of hydrolysis degree of proteins. Analytical Methods, 9(25), 3937–3945. https://doi.org/10.1039/C7AY00865A
32. Zhang, Z., Adebayo, I. A., O'Shea, R., & Astatkie, T. (2024). Production of a high-quality peptone from black soldier fly (Hermetia illucens L.) larvae defatted meal using enzymatic hydrolysis. Heliyon, 10(7), e28664. https://doi.org/10.1016/j.heliyon.2024.e28664