journal article·2025-11-20Open access

Comparison of Pretreatment Methods for Obtaining Collagen Hydrolysates from the Swim Bladder of Totoaba macdonaldi and Their Negative Impact on Cancer Cells

Evelin Martínez-Benavidez, Angélica María Vergara-Pineda, Jesús Cervantes-Martínez, José Leonardo Puch-Sánchez, Sandra Daniela Bravo, Ofelia Yadira Lugo-Melchor, Hugo S. García, Inocencio Higuera-Ciapara

Applied Sciences· Volume 15· Issue 22

Abstract

The search for therapeutic bioactive peptides has led to the utilization of marine byproducts as collagen sources. This study evaluated the effect of collagen hydrolysates (CH) obtained from the swim bladder (SB) of Totoaba macdonaldi on breast (MCF-7) and colorectal (Caco-2) adenocarcinoma cells and on human dermal fibroblasts (CRL-1474), considering the need for less invasive and less toxic treatment alternatives. Two pretreatment methods for the SB were compared: (1) NaOH and butanol (SBPT), and (2) hexane (SBDF). The pretreated tissues underwent direct enzymatic hydrolysis using bromelain. The resulting hydrolysates were characterized by SDS-PAGE, Raman spectroscopy, and chromatographic profiling. Both pretreatments preserved the structure of type I collagen. Bromelain hydrolysis was efficient, yielding peptides with molecular weights below 20 kDa for CH-SBPT and below 10 kDa for CH-SBDF. CH of Totoaba macdonaldi significantly reduced MCF-7 and Caco-2 cells viability, particularly at 20 mg/mL. In CRL-1474 fibroblasts, CH-SBDF stimulated cell proliferation, while CH-SBPT had neutral effects. Hexane pretreatment is a viable alternative to NaOH, reducing processing steps without compromising yield or bioactivity. CH derived from Totoaba macdonaldi exhibit promising anticancer and regenerative properties, suggesting potential biomedical applications. Further research is needed to isolate specifically active peptides and elucidate their mechanisms of action.
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Research context

OpenAlex topics: Protein Hydrolysis and Bioactive Peptides · Seaweed-derived Bioactive Compounds · Collagen: Extraction and Characterization

SciVX discovery tags are terminology-based. OpenAlex topics are source-provided classifications.

Affiliations

Unidad de Servicios Analíticos y Metrológicos, Centro de Investigación y Asistencia en Tecnología y Diseño del Estado de Jalisco, A. C. (CIATEJ), Guadalajara 44270, Jalisco, Mexico

Unidad de Investigación y Desarrollo en Alimentos, Tecnológico Nacional de México Campus Veracruz, Veracruz 91897, Veracruz, Mexico

Centro de Investigación en Alimentación y Desarrollo, A.C., Hermosillo 83304, Sonora, Mexico

Dirección de Investigación y Desarrollo, Universidad Anáhuac Mayab, S.C., Mérida 97302, Yucatán, Mexico

References (39)

Show source-provided references
1. Collagen as a source of bioactive peptides: A bioinformatics approach10.1016/j.ejbt.2020.09.009 ↗
2. The wound healing potential of collagen peptides derived from the jellyfish Rhopilema esculemtum10.1016/j.cjtee.2018.10.004 ↗
3. Vieira, H., Lestre, G.M., Solstad, R.G., Cabral, A.E., Botelho, A., Helbig, C., Coppola, D., De Pascale, D., Robbens, J., and Raes, K. (2023). Current and expected trends for the marine chitin/chitosan and collagen value chains. Mar. Drugs, 21.10.20944/preprints202310.1827.v1 ↗
4. (2024, December 02). CITES. Available online: https://cites.org/eng/node/48556.
5. Coscueta, E.R., Brassesco, M.E., and Pintado, M. (2021). Collagen-Based bioactive bromelain hydrolysate from Salt-Cured cod Skin. Appl. Sci., 11.10.3390/app11188538 ↗
6. Bioactive food-derived peptides for functional nutrition: Effect of fortification, processing and storage on peptide stability and bioactivity within food matrices10.1016/j.foodchem.2022.135046 ↗
7. Nong, N.T.P., and Hsu, J.L. (2022). Bioactive peptides: An understanding from current screening methodology. Processes, 10.10.3390/pr10061114 ↗
8. Zaky, A.A., Simal-Gandara, J., Eun, J.B., Shim, J.H., and Abd El-Aty, A.M. (2022). Bioactivities, applications, safety, and health benefits of bioactive peptides from food and by-products. A review. Front. Nutr., 8.10.3389/fnut.2021.815640 ↗
9. Marine bioactive peptides with anticancer potential, a narrative review10.62347/TUVQ7468 ↗
10. Antiproliferative activity of fish protein hydrolysates on human breast cancer cell lines10.1016/j.procbio.2005.11.024 ↗
11. Evaluation of the efficacy of proline rich VPPPVPPRRR Peptide (VPP) in MCF-7 cells10.1007/s10989-025-10706-1 ↗
12. Taroncher, M., Rodríguez-Carrasco, Y., Aspevik, T., Kousoulaki, K., Barba, J., and Ruiz, M.-J. (2021). Cytoprotective effects of fisch protein hydrolysates against H2O2-induced oxidatuve stress and mycotoxins in Caco-2/TC7 cells. Antioxidants, 10.10.3390/antiox10060975 ↗
13. Karcioglu, B.L., Yavas, C., and Hekim, N. (2025). Kiperin double-hydrolyzed collagen as potential anti-tumor agent: Effects on HCT116 colon carcinoma cells and oxidative stress modulation. Curr. Issues Mol. Biol., 47.10.3390/cimb47050364 ↗
14. Hernández-Ledesma, B., and Chia, C.H. (2013). Bioactive Food Peptides in Health and Disease, IntechOpen.10.5772/3318 ↗
15. Peptides from bovine bone gelatin hydrolysate with anticancer activity against the human ovary carcinoma cell line and its possible mechanism10.1016/j.jff.2024.106061 ↗
16. Collagen peptides modulate the metabolism of extracellular matrix by human dermal fibroblasts derived from sun-protected and sun-exposed body sites10.1002/cbin.10872 ↗
17. Food-derived bioactive peptides: Production, biological activities, opportunities and challenges10.1016/j.jfutfo.2022.08.002 ↗
18. (2017). Oficio No. SGPA/DIGIRA/DG/06043 (Standard No. DGVS-UMA-IN-1821-SON/17).
19. Cruz-López, H., Rodríguez Morales, S., Enríquez Paredes, L.M., Villareal Gómez, L.J., True, C., Olivera Castillo, L., Fernández Velasco, D.A., and López, L.M. (2023). Swin bladder of Totoaba macdonaldi: A source of value-added collagen. Mar. Drugs, 21.10.3390/md21030173 ↗
20. Cleavage of structural proteins during the assembly of the head of bacteriophage T410.1038/227680a0 ↗
21. Rapid colorimetric assay for cellular growth and survival: Application to proliferation and cytotoxicity assays10.1016/0022-1759(83)90303-4 ↗
22. Marine origin collagens and its potential applications10.3390/md12125881 ↗
23. Structural, physicochemical properties and function of swim bladder collagen in promoting fibroblasts viability and collagen synthesis10.1016/j.lwt.2022.114294 ↗
24. Sun, Y., Cui, Y., Wang, R., Ma, J., Sun, H., Chen, L., and Yang, R. (2023). The hydrolysis of pigment-protein phycoerythrin by bromelain enhances the color stability. Foods, 12.10.3390/foods12132574 ↗
25. Bromelain: A Potent Phytomedicine
26. Optimization of collagen extraction from chicken feet by papain hydrolysis and synthesis of chicken feet collagen based biopolymeric fibres10.1016/j.fbio.2018.03.003 ↗
27. Hydrolyzed collagen: Exploring its applications in the food and beverage industries and assessing its impact on human health—A comprehensive review10.1016/j.heliyon.2024.e36433 ↗
28. Li, Q., Chang, B., Huang, G., Wang, D., Gao, Y., Fan, Z., Sun, H., and Sui, X. (2025). Differential Enzymatic Hydrolysis: A Study on its impact on soy protein structure, function, and soy milk powder properties. Foods, 14.10.3390/foods14050906 ↗
29. Synytsya, A., Janstová, D., Smidová, M., Synytsya, A., and Petrtyl, J. (2023). Evaluation of IR and Raman spectroscopic markers of human collagens: Insides for indicating colorectal carcinogenesis. Spectrochim. Acta A Mol. Biomol. Spectrosc., 296.10.1016/j.saa.2023.122664 ↗
30. Khalid, M., Bora, T., Ghaithi, A.A., Thukral, S., and Dutta, J. (2018). Raman Spectroscopy detects changes in bone mineral quality and collagen cross-linkage in Staphylococcus infected human bone. Sci. Rep., 8.10.1038/s41598-018-27752-z ↗
31. Bak, S.Y., Lee, S.W., Choi, C.H., and Kim, H.W. (2018). Assessment of the influence of acetic acid residue on type I collagen during isolation and characterization. Materials, 11.10.3390/ma11122518 ↗
32. Połomska, M., Kubisz, L., Wolak, J., and Hojan-Jezierska, D. (2021). Effects of Temperature on the FT NIR Raman Spectra of Fish Skin Collagen. Appl. Sci., 11.10.3390/app11188358 ↗
33. Anticancer potential of bioactive peptides from animal sources (Review)10.3892/or.2017.5778 ↗
34. Induction of apoptosis in a human lymphoma cell line by hydrophobic peptide fraction separated from anchovy sauce10.1002/biof.552210112 ↗
35. A fish antimicrobial peptide, tilapia hepcidin TH2-3, shows potent antitumor activity against human fibrosarcoma cells10.1016/j.peptides.2009.06.009 ↗
36. Review on marine collagen peptides induce cancer cell apoptosis, necrosis, and autophagy by reducing oxidized free radicals10.32604/biocell.2023.027729 ↗
37. Dierckx, S., Patrizi, M., Merino, M.a., González, S., Mullor, J.L., and Nergiz-Unal, R. (2024). Collagen peptides affect collagen synthesis and the expression of collagen, elastin, and versican genes in cultured human dermal fibroblasts. Front. Med., 11.10.3389/fmed.2024.1397517 ↗
38. Hu, Z., Yang, P., Zhou, C., Li, S., and Hong, P. (2017). Marine collagen peptides from the skin of Nile tilapia (Oreochromis niloticus): Characterization and wound healing evaluation. Mar. Drugs, 15.10.3390/md15040102 ↗
39. Shahidi, F., and Saeid, A. (2025). Bioactivity of marine-derived peptides and proteins: A review. Mar. Drugs, 23.10.3390/md23040157 ↗