Extraction of acid-soluble and pepsin-soluble collagens from common carp scales (Cyprinus carpio) and determination of their characteristics

Document Type : Full Research Paper

Authors

• Marjan Zargar ¹
• Bahareh Shabanpour ¹
• Parastoo Pourashouri ¹
• Ebrahim Zabihi Neyshbouri ²

¹ Fisheries Products Processing Department, Gorgan University of Agricultural Sciences and Natural Resources, Gorgan, Iran.

² Department of Pharmacology, Babol University of Medical Sciences, Iran.

https://doi.org/10.22067/ifstrj.2022.74164.1121

Abstract

[1]Introduction: Collagen is the most abundant and important structural protein in the connective tissue of animals, the production of which is of great importance in the fields of medicine, cosmetics and food. Due to religious restrictions as well as common diseases between livestock and humans, today collagen extraction has turned to other sources, especially aquatic sources. Therefore, the aim of this study was to extract collagen by conventional acidic and enzymatic methods from common carp scales and determine its characteristics in order to make optimal use of this waste to produce valuable products and find alternatives to collagen obtained from land animals.
 
Material and Method: Acid-soluble collagen (ASC) and pepsin-soluble collagen (PSC) were extracted from common carp scales (Cyprinus carpio) and their properties were determined. Common carp scales were prepared with the use of 0.5 M acetic acid and pepsin enzyme. Finally, the extracted collagens were lyophilized and after calculating the extraction efficiencies, their characteristics were determined by electrophoresis tests, UV spectroscopy, X-ray diffraction and isoelectric point determination.
 
 
Results and Discussion: The results showed that the extraction efficiencies of ASC and PSC were 1.9% and 2.96% (based on dry weight), respectively, which means digestion with pepsin could increase collagen efficiency up to 1.54 times. The results of SDS-PAGE analysis showed that both ASC and PSC are type I collagen and are composed of α₁, α₂ and β in the (α₁) ₂α₂ chain structure; the isoelectric point of collagens was in the pH range of 5-6. The maximum absorption peak of the UV spectrum of collagen was observed at 235 nm. Although pepsin enzyme (1% dry weight of scales) increases efficiency without significant changes in collagen native structure, but its use for mass production of type I collagen in Iran is not recommended unless self-sufficiency and reduction of pepsin price achieved, while collagen extraction by acidic method is very simple and research to design a production line for this method is recommended. Common carp scales have the potential to be used as an alternative source of collagen in the food and health-pharmaceutical industries. These results can provide a solution to control the waste of the aquatic processing industry in creating environmental pollution, as well as producing a high value-added product from common carp scales.

Keywords

• Scales
• Common carp (Cyprinus carpio)
• Acid-soluble collagen
• Pepsin-soluble collagen
• Waste

Main Subjects

• Food Chemistry

References

1. Ali AMM, Benjakul S, Prodpran T and Kishimura H, (2018). Extraction and characterisation of collagen from the skin of golden carp (Probarbus jullieni), a processing by-product. Waste Biomass Valori 9: 783-791. https://doi.org/10.1007/s12649-017-9841-0
2. Ali AMM, Hideki K and Soottawat B, (2018). Extraction efficiency and characteristics of acid and pepsin soluble collagens from the skin of golden carp (Probarbus jullieni) as affected by ultrasonication. Process Biochemistry, 66: 237-244. https://doi.org/10.1016/j.procbio.2018.01.003
3. Alves AL, Marques ALP, Martins E, Silva TH and Reis RL, (2017). Cosmetic potential of marine fish skin collagen. Cosmetics 4: 39. https://doi.org/10.3390/cosmetics4040039
4. Barzideh, Z, Latiff AA, Gan CY, Benjakul S and Karim AA, (2013). Isolation and characterisation of collagen from the ribbon jellyfish (Chrysaora sp.). International Journal of Food Science and Technology.  49(6), 1490-1499. https://doi.org/10.1111/ijfs.12464
5. Benjakul S, Thiansilakul Y, Visessanguan W, Roytrakul S, Kishimura H and Prodpran T, (2010). Extraction and characterisation of pepsin solubilised collagens from the skin of bigeye snapper (Priacanthus tayenus and Priacanthus macracanthus). Journal of the Science Food and Agriculture 90(1): 132-138. https://doi.org/10.1002/jsfa.3795
6. Chai HJ, Li JH, Huang HN, Li TL, Chan YL, Shiau CY and Wu CJ, (2010). Effects of sizes and conformations of fish-scale collagen peptides on facial skin qualities and transdermal penetration efficiency. Journal of Biomedicine and Biotechnology. https://doi.org/10.1155/2010/757301
7. Chattopadhyay S and Raines RT, (2014). Collagen-Based Biomaterials for Wound Healing. Biopolymers 101(8): 821-833. https://doi.org/10.1002/bip.22486
8. Chen Z, Wei B, Mo X, Lim CT, Ramakrishna S and Cui F, (2009). Mechanical properties of electro spun collagen–chitosan complex single fibers and membrane. Materials Science and Engineering C 29 (8): 2428-2435. https://doi.org/10.1016/j.msec.2009.07.006
9. Chinh NT, Manh VQ, Trung VQ, Trang TDM, Hoang T, (2019). Characterization of collagen derived from tropical freshwater carp fish scale wastes and its amino acid sequence. Natural Product Communications 14(7).
10. Dermal Fillers, (2013). The Ageing Skin. Pharmaxchange. info. Retrieved on 2013-04-21.
11. El-Rashidy AA, Gad A, Abu-Hussein AE, Habib S, Badr NA and Hashem AA, (2015). Chemical and biological evaluation of Egyptian Nile Tilapia (Oreochromis niloticas) fish scale collagen. International Journal of Biological Macromolecules 79: 618-626. https://doi.org/10.1016/j.ijbiomac.2015.05.019
12. Hashim P, Mohd Ridzwan MS, Bakar J and Hashim DM, (2015). Collagen in food and beverage industries. International Food Research Journal 22 (1): 1-8.
13. Hong H, Roy BC, Chalamaiah M, Bruce HL and Wu JP, (2018). Pretreatment with formic acid enhances the production of small peptides from highly cross-linked collagen of spent hens. Food Chemistry 258: 174. https://doi.org/10.1016/j.foodchem.2018.03.036
14. Khong NMH, Yusoff FMd, Jamilah B, Basria M, Maznaha I, Chana KW, Armaniaa N and Nishikawad J, (2018). Improved collagen extraction from jellyfish (Acromitus hardenbergi) with increased physical-induced solubilization processes. Food Chemistry 251: 41-50. https://doi.org/10.1016/j.foodchem.2017.12.083
15. Krishnan S and Perumal P, (2013). Preparation and biomedical characterization of jellyfish (Chrysaora quinquecirrha) collagen from southest coast of India. International Journal of Pharmacy and Pharmaceutical Sciences 5 (3).
16. Laemmli UK, (1970). Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227: 680-685. https://doi.org/10.1038/227680a0
17. Li LY, Zhao YQ, He Y, Chi CF and Wang B, (2018). Physicochemical and antioxidant properties of acidand pepsin-soluble collagens from the scales of miiuy croaker (Miichthys miiuy). Marine Drugs 16: 394. https://doi.org/10.3390/md16100394
18. Lim YS, Ok YJ, Hwang SY, Kwak JY and Yoon S, (2019). Marine collagen as a promising biomaterial for biomedical applications. Marine Drugs 17: 467. https://doi.org/10.3390/md17080467
19. Matmaroh K, Benjakul S, Prodpran T, Encarnacion AB and Kishimura H, (2011). Characteristics of acid soluble collagen and pepsin soluble collagen from scale of spotted golden goatfish (Parupeneus heptacanthus). Food Chemistry 129(3): 1179-1186. https://doi.org/10.1016/j.foodchem.2011.05.099
20. Miller EJ and Matukas VJ, (1969). Chick cartilage collagen: A new type of a1 chain not present in bone or skin of the species. Proceedings of the National Academy of Sciences of the United States of America 64: 1264-1268. https://doi.org/10.1073/pnas.64.4.1264
21. Nagai T, Izumi M and Ishii M, (2004). Fish scale collagen. Preparation and partial characterization. International Journal of Food Science and Technology 39(3): 239-244. https://doi.org/10.1111/j.1365-2621.2004.00777.xNomura y and Kitozome N, (2002). Use of shark collagen for cell culture and zymography. Bioscience Biotechnology, Biochemistry 66 (12): 2673-2676. https://doi.org/10.1271/bbb.66.2673
22. Pal P, Srivas PK, DadhichP, Das B, Maity PP, Moulik D and Dhara S, (2016). Accelerating full thickness wound healing using collagen sponge of mrigal fish (Cirrhinus cirrhosus) scale origin. International Journal of Biological Macromolecules 93: 1507-1518. https://doi.org/10.1016/j.ijbiomac.2016.04.032
23. Panda NN, Pramanik K and Sukla LB, (2014). Extraction and characterization of biocompatible hydroxyapatite from fresh water fish scales for tissue engineering scaffold. Bioprocess Biosystem 37: 433. https://doi.org/10.1007/s00449-013-1009-0
24. Pati F, Adhikari B and Dhara S, (2010). Isolation and characterization of fish scale collagen of higher thermal stability. Bioresource Technology 101(10): 3737-3742. https://doi.org/10.1016/j.biortech.2009.12.133
25. Ramshaw JA, Peng Y, Glattauer V and Werkmeister JA, (2009). Collagens as biomaterials. Journal of Materials Science: Materials in Medicine 20 (1): S3-S8. https://doi.org/10.1007/s10856-008-3415-4
26. Richard-Blum S, (2011). Collagen family. Cold Spring Harbor perspectives in biology 3: a004978. DOI: 1101/cshperspect.a004978
27. Slimane EB and Sadok S, (2018). Collagen from cartilaginous fish by-products for a potential application in bioactive film composite. Marine Drugs 16: 211. https://doi.org/10.3390/md16060211
28. Said MI, (2020). IOP Conference Series: Earth and Environronmental Science. 492, 012086. DOI:10.1088/1755-1315/492/1/012086
29. Wu J, Kong L, Zhang J and Chen W, (2019). Extraction and properties of acid-soluble collagen and pepsin-soluble collagen from silver carp (Hypophthalmichthys molitrix) Scales: Prerequisite information for fishery processing waste reuse. Polish Journal of Environmental Studies 28(4): 2923-2930. https://doi.org/10.1007/s10068-015-0264-z
30. Wu QQ, Li T, Wang B, Ding GF, (2015). Preparation and characterization of acid and pepsin-soluble collagens from scales of croceine and redlip croakers. Food Science Biotechnology 24: 2003-2010. https://doi.org/10.1007/s10068-015-0264-z
31. Shamansky LM, Pratt D, Boisvenue RJ and Cox GN, (1989). Cuticle collagen genes of haemonchus-contortus and caenorhabditis-elegans are highly conserved. Molecular and Biochemical Parasitology 37 (1): 73. https://doi.org/10.1016/0166-6851(89)90104-7
32. Yu D, Chi CF, Wang B, Ding GF and Li Z, 2014. Characterization of acid and pepsin soluble collagens from spine and skull of skipjack tuna (Katsuwonus pelamis). Chinese Journal of Natural Medicines 12: 712-720.
33. Zhang J, Duan R, Ye C and Konno K, 2010. Isolation and characterization of collagens from scale of silver carp (Hypophthalmichthys molitrix). Journal of Food Biochemistry 34(6): 1343-1354. https://doi.org/10.1111/j.1745-4514.2010.00439.x
34. Zhang F, Wang A, Li Z, He S and Shao L, (2011). Preparation and characterisation of collagen from freshwater fish scales. Food Nutrient Science 2: 818-823. DOI:10.4236/fns.2011.28112