Enzymatic extraction of sulfated polysaccharide from the skin of rainbow trout (Oncorhynchus mykiss) and evaluation of its chemical, antioxidant and functional properties

مجله پژوهشهای علوم و صنایع غذایی ایران

Volume 18, Issue 5 - Serial Number 77, November and December 2022, Pages 725-738   

Document Type: Full Research Paper

DOI: 10.22067/ifstrj.2022.72999.1102

Authors

Shahab Naghdi¹; Masoud Rezaei^{* 1}; Mehdi Abdollahi²; Mehdi Tabarsa³

¹Department of Fishery Processing, Faculty of Natural Resources and Marine Sciences, Tarbiat Modares University, Noor, Iran.

²Department of Biology and Biological Engineering, Food Science and Nutrition, Chalmers University of Technology, Gothenburg, Sweden.

³Department of Fishery Products Processing, Faculty of Natural Resources and Marine Sciences, Tarbiat Modares University, Noor, Iran

Abstract

[1]Introduction: Bioactive compounds are substances found in small amounts in food. In addition to their influence on human development, these compounds also play a crucial role in reducing diseases in human. Polysaccharides are a group of bioactive compounds that come from a variety of sources. Polysaccharides are macromolecules that are usually composed of more than ten monosaccharides. The constituent monosaccharides are arranged linearly or branched together through glycoside bonds, depending on the length of the chain and the number of constituent monosaccharides. They also have different molecular weight. Polysaccharides, like other essential macromolecules such as proteins and poly-nucleotides in the body, are essential for the flaxseed body's daily activities and play an influential role in cell-cell communication, cell adhesion, and the identification of molecules in the immune system. A group of polysaccharides derived from marine sources are sulfated polysaccharides. These polysaccharides are a broad branch of the resulting polysaccharides. In industrial quantities, sulfated polysaccharides are produced from pig skin and pig bone, and there are some restrictions on the use of these products in some countries. The limitations on the use of these products made from pig waste are the risks of transmitting influenza, as well as the prohibition of pork in some Islāmic countries. In this regard, by-products from seafood processing, which account for about 20 to 50 percent of the initial weight of raw material, are one of the sources that researchers are considering to extract these compounds.
 
Material and Method: After preparation of the by-product, it was covered with ice in a ratio of 1 to 3 and transferred to the laboratory of Tarbiat Modares University. The sample was then washed and then ground. Finally, it was packed in plastic bags and kept in the freezer at -18 ° C until the day of experiment. Then, the enzymatic hydrolysis method and precipitation by ethanol were used to get sulfated polysaccharides. Chemical analyses were performed to determine carbohydrates, sulfates, proteins, and uronic acid content. The FTIR spectrum of extracted sulfate polysaccharide was determined using an FTIR spectrophotometer in the range of 400-4000 cm^-1. Evaluation of antioxidant properties of obtained sulfate polysaccharide was assessed by DPPH free radical scavenging activity, ABTS free radical scavenging activity, and ferrozine tests. Emulsifying and foaming properties were also evaluated as functional properties.
 
Results and Discussion: In the present study, sulfated polysaccharide was extracted from Rainbow trout (Oncorhynchus mykiss) skin by pepsin enzyme and its FTIR spectrum, carbohydrate, sulfate, uronic acid and protein were analyzed. The results of the chemical analysis of the extracted polysaccharide showed that the extraction efficiency was 3.23± 0.02%, as well as the percentage of carbohydrate and protein of the obtained polysaccharide was 57.03± 2 2.56, 7.78± 0.43% respectively. Also, the amount of sulfate and uronic acid were 6.54± 0.77 and 3.86± 0.43, respectively. The results of infrared spectroscopy showed the presence of a broad peak in the range between 3350 and 3450 cm^-1 for the –OH group and the S=O sulfate flexural band in the range of 1245 cm^-1. An increasing and significant trend was observed in different concentrations used for the DPPH test (p <0.05) which had the highest neutralizing power (38.85%) at a concentration of 2 mg/ml. The highest percentage of ABTS radical chelating was observed at a concentration of 4 mg/mm of distilled water with 71.70% (p <0.05). The chelating results of the extracted polysaccharide against ferrous ions showed that the highest chelating percentage was 98.43% (p <0.05). The foaming capacity, stability properties of the foam, and the emulsifying ability of the studied sample showed a trend of increasing the concentration coefficient of the sample (p <0.05), and the concentration of 10% used sulfated polysaccharide had the highest foaming percentage (72/22%) and foam stability (62.22%) (p <0.05). The emulsifying property of extracted sulfate polysaccharide against soybean oil was higher in all concentrations used than sunflower oil (p <0.05), and the highest value of that was related to the concentration of 10% with 86.57% and 92.59% against sunflower oil and soybean oil (p <0.05). The obtained results demonstrated that the fish skin extracted polysaccharide can serve as a natural antioxidant and functional agent in the food industry                           

Keywords

Rainbow trout (Oncorhynchus mykiss); Sulfated polysaccharide; Antioxidant properties; Foaming properties; Emulsifying properties

References

1. Alboofetileh, M., Rezaei, M., & Tabarsa, M. (2018). Enzyme-assisted extraction of Nizamuddinia zanardinii for the recovery of sulfated polysaccharides with anticancer and immune-enhancing activities. Journal of Applied Phycology. https://doi.org/10.1007/s10811-018-1651-7
2. Alboofetileh, M., Rezaei, M., Tabarsa, M., & You, S. (2019). Bioactivities of Nizamuddinia zanardinii sulfated polysaccharides extracted by enzyme, ultrasound and enzyme-ultrasound methods. Journal of Food Science and Technology. https://doi.org/10.1007/s13197-019-03584-1
3. Arima, K., Fujita, H., Toita, R., Imazu-Okada, A., Tsutsumishita-Nakai, N., Takeda, N., Nakao, Y., Wang, H., Kawano, M., Matsushita, K., Tanaka, H., Morimoto, S., Nakamura, A., Kitagaki, M., Hieda, Y., Hatto, R., Watanabe, A., Yumura, T., Okuhara, T., … Tamura, J. I. (2013). Amounts and compositional analysis of glycosaminoglycans in the tissue of fish. Carbohydrate Research, 366, 25–32. https://doi.org/10.1016/j.carres.2012.11.010
4. Bitter, T., & Muir, H. M. (1962). A modified uronic acid carbazole reaction. In Analytical Biochemistry (Vol. 4, Issue 4, pp. 330–334). https://doi.org/10.1016/0003-2697(62)90095-7
5. Borazjani, N. J., Tabarsa, M., You, S. G., & Rezaei, M. (2017). Effects of extraction methods on molecular characteristics, antioxidant properties and immunomodulation of alginates from Sargassum angustifolium. International Journal of Biological Macromolecules, 101, 703–711. https://doi.org/10.1016/j.ijbiomac.2017.03.128
6. Chang, S. C., Hsu, B. Y., & Chen, B. H. (2010). Structural characterization of polysaccharides from Zizyphus jujuba and evaluation of antioxidant activity. International Journal of Biological Macromolecules, 47(4), 445–453. https://doi.org/10.1016/j.ijbiomac.2010.06.010
7. Chen, G., Fang, C., Chen, X., Wang, Z., Liu, M., & Kan, J. (2019). High-pressure ultrasonic-assisted extraction of polysaccharides from Mentha haplocalyx: Structure, functional and biological activities. Industrial Crops and Products, 130(October 2018), 273–284. https://doi.org/10.1016/j.indcrop.2018.12.086
8. Chew, K. K., Ng, S. Y., Thoo, Y. Y., Khoo, M. Z., Wan Aida, W. M., & Ho, C. W. (2011). Effect of ethanol concentration, extraction time and extraction temperature on the recovery of phenolic compounds and antioxidant capacity of Centella asiatica International Food Research Journal, 18(2), 571–578.
9. Cho, C. W., Han, C. ji, Rhee, Y. K., Lee, Y. C., Shin, K. S., Shin, J. S., Lee, K. T., & Hong, H. Do. (2015). Cheonggukjang polysaccharides enhance immune activities and prevent cyclophosphamide-induced immunosuppression. International Journal of Biological Macromolecules, 72, 519–525. https://doi.org/10.1016/j.ijbiomac.2014.09.010
10. Dong, X., Duan, X., Sun, Z., Zhang, X., Li, C., Yang, S., Ren, B., Zheng, S., & Dionysiou, D. D. (2019). Structural characterization and antioxidant activities of a water soluble polysaccharide isolated from Glycyrrhiza glabra. “Applied Catalysis B, Environmental,” 118214. https://doi.org/10.1016/j.apcatb.2019.118214
11. Dubois, M., Gilles, K. A., Hamilton, J. K., Rebers, P. A., & Smith, F. (1956). Colorimetric Method for Determination of Sugars and Related Substances. Analytical Chemistry, 28(3), 350–356. https://doi.org/10.1021/ac60111a017
12. FAO (2018), Yearbook of Fishery Statistics, Statistics and Information Service, FAO Fisheries and Aquaculture Department. Viale delle Terme di Caracalla 00153, Rome: Food and Aquaculture Organization of the United Nations. http://www.fao.org/fishery/statistics/en.
13. Ghribi, A. M., Sila, A., Gafsi, I. M., Blecker, C., Danthine, S., Attia, H., ... & Besbes, S. (2015). Structural, functional, and ACE inhibitory properties of water-soluble polysaccharides from chickpea flours. International Journal of Biological Macromolecules, 75, 276-282. https://doi.org/10.1016/j.ijbiomac.2015.01.037
14. Gilar, M., Yu, Y. Q., Ahn, J., Xie, H., Han, H., Ying, W., & Qian, X. (2011). Characterization of glycoprotein digests with hydrophilic interaction chromatography and mass spectrometry. Analytical Biochemistry, 417(1), 80–88. https://doi.org/10.1016/j.ab.2011.05.028
15. Gomaa, M., Hifney, A. F., Fawzy, M. A., & Abdel-Gawad, K. M. (2018). Use of seaweed and filamentous fungus derived polysaccharides in the development of alginate-chitosan edible films containing fucoidan: Study of moisture sorption, polyphenol release and antioxidant properties. In Food Hydrocolloids (Vol. 82). Elsevier Ltd. https://doi.org/10.1016/j.foodhyd.2018.03.056 
16. Grina, F., Ullah, Z., Kaplaner, E., Moujahid, A., Eddoha, R., Nasser, B., Terzioğlu, P., Yilmaz, M. A., Ertaş, A., Öztürk, M., & Essamadi, A. (2020). In vitro enzyme inhibitory properties, antioxidant activities, and phytochemical fingerprints of five Moroccan seaweeds. South African Journal of Botany, 128, 152–160. https://doi.org/10.1016/j.sajb.2019.10.021 
17. Guo, L., Zhu, W., Xu, F., Liu, M., Xie, Y., & Zhang, J. (2014). Optimized ultrasonic-assisted extraction of polysaccharides from Cyclina sinensis and evaluation of antioxidant activities in vitro. CYTA- Journal of Food, 12(1), 32–39. https://doi.org/10.1080/19476337.2013.785982 
18. Hamzaoui, A., Ghariani, M., Sellem, I., Hamdi, M., Feki, A., Jaballi, I., Nasri, M., & Amara, I. Ben. (2020). Extraction, characterization and biological properties of polysaccharide derived from green seaweed “Chaetomorpha linum” and its potential application in Tunisian beef sausages. International Journal of Biological Macromolecules, 148, 1156–1168. https://doi.org/10.1016/j.ijbiomac.2020.01.009
19. Joana Gil-Chávez, G., Villa, J. A., Fernando Ayala-Zavala, J., Basilio Heredia, J., Sepulveda, D., Yahia, E. M., & González-Aguilar, G. A. (2013). Technologies for extraction and production of bioactive compounds to be used as nutraceuticals and food ingredients: An Overview. Comprehensive Reviews in Food Science and Food Safety, 12(1), 5–23. https://doi.org/10.1111/1541-4337.12005
20. Jridi, M., Mezhoudi, M., Abdelhedi, O., Boughriba, S., Elfalleh, W., Souissi, N., Nasri, R., & Nasri, M. (2018). Bioactive potential and structural characterization of sulfated polysaccharides from Bullet tuna (Auxis Rochei) by-products. Carbohydrate Polymers, 194(April), 319–327. https://doi.org/10.1016/j.carbpol.2018.04.038
21. Jridi, M., Nasri, R., Marzougui, Z., Abdelhedi, O., Hamdi, M., & Nasri, M. (2019). Characterization and assessment of antioxidant and antibacterial activities of sulfated polysaccharides extracted from cuttlefish skin and muscle. International Journal of Biological Macromolecules, 123, 1221–1228. https://doi.org/10.1016/j.ijbiomac.2018.11.170
22. Khan, B. M., Qiu, H. M., Wang, X. F., Liu, Z. Y., Zhang, J. Y., Guo, Y. J., Chen, W. Z., Liu, Y., & Cheong, K. L. (2019). Physicochemical characterization of Gracilaria chouae sulfated polysaccharides and their antioxidant potential. International Journal of Biological Macromolecules, 134, 255–261. https://doi.org/10.1016/j.ijbiomac.2019.05.055
23. Leroux, J., Langendorff, V., Schick, G., Vaishnav, V., & Mazoyer, J. (2003). Emulsion stabilizing properties of pectin. Food Hydrocolloids, 17(4), 455–462. https://doi.org/10.1016/S0268-005X(03)00027-4
24. Li, H., Tao, Y., Zhao, P., Zhi, D., Gao, X., Zhao, X., & Li, M. (2019). Effect of ultrasound-assisted extraction on physicochemical properties and TLR2-affinity binding of the polysaccharides from Pholiota nameko. International Journal of Biological Macromolecules, 135, 1020–1027. https://doi.org/10.1016/j.ijbiomac.2019.05.177
25. Li, X., & Wang, L. (2016). Effect of extraction method on structure and antioxidant activity of Hohenbuehelia serotina polysaccharides. International Journal of Biological Macromolecules, 83, 270–276. https://doi.org/10.1016/j.ijbiomac.2015.11.060
26. Lowry, O. H., Rosebrough, N. J., Farr, A. L., & Randall, R. J. (1951). Protein measurement with the Folin phenol reagent. The Journal of Biological Chemistry, 193(1), 265–275. https://doi.org/10.1016/0304-3894(92)87011-4
27. Loyd, A. G., Dogson, K. S., Price, R. G., & Rose, F. A. (1960). Polysaccharide Sulphates. Biochem Biophys Acta, 46(2961).
28. Odeleye, T., White, W. L., & Lu, J. (2019). Extraction techniques and potential health benefits of bioactive compounds from marine molluscs: a review. Food & Function. https://doi.org/10.1039/c9fo00172g
29. Bhuyar, M.H. Rahim, S. Sundararaju, G.P. Maniam, N. G. (2020). Antioxidant and antibacterial activity of red seaweed; Kappaphycus alvarezii against pathogenic bacteria. Global Journal of Environmental Science and Management, 5(1), 47–58. https://doi.org/10.22034/gjesm.2020.01.0
30. Pan, X. Y., Wang, Y. M., Li, L., Chi, C. F., & Wang, B. (2019). Four antioxidant peptides from protein hydrolysate of red stingray (dasyatis akajei) cartilages: Isolation, identification, and in vitro activity evaluation. Marine Drugs, 17(5). https://doi.org/10.3390/md17050263
31. Pomin, V. H., & Mulloy, B. (2018). Glycosaminoglycans and proteoglycans. Pharmaceuticals, 11(1), 1–9. https://doi.org/10.3390/ph11010027
32. Qi, H., Zhao, T., Zhang, Q., Li, Z., Zhao, Z., & Xing, R. (2005). Antioxidant activity of different molecular weight sulfated polysaccharides from Ulva pertusa Kjellm (Chlorophyta). Journal of Applied Phycology, 17(6), 527–534. https://doi.org/10.1007/s10811-005-9003-9
33. Rjeibi, I., Hentati, F., Feriani, A., Hfaiedh, N., Delattre, C., Michaud, P., & Pierre, G. (2019). Novel antioxidant, anti-α-amylase, anti-inflammatory and antinociceptive water-soluble polysaccharides from the aerial part of Nitraria retusa. Foods, 9(1), 28. https://doi.org/10.3390/foods9010028
34. Romdhane, M. Ben, Haddar, A., Ghazala, I., Jeddou, K. Ben, Helbert, C. B., & Ellouz-Chaabouni, S. (2017). Optimization of polysaccharides extraction from watermelon rinds: Structure, functional and biological activities. Food Chemistry, 216, 355–364. https://doi.org/10.1016/j.foodchem.2016.08.056
35. Saravana, P. S., Cho, Y. J., Park, Y. B., Woo, H. C., & Chun, B. S. (2016). Structural, antioxidant, and emulsifying activities of fucoidan from Saccharina japonica using pressurized liquid extraction. Carbohydrate Polymers, 153, 518–525. https://doi.org/10.1016/j.carbpol.2016.08.014
36. Sciarini, L. S., Maldonado, F., Ribotta, P. D., Pérez, G. T., & León, A. E. (2009). Chemical composition and functional properties of Gleditsia triacanthos Food Hydrocolloids, 23(2), 306–313. https://doi.org/10.1016/j.foodhyd.2008.02.011
37. Shahidi, F. (2012). Nutraceuticals, functional foods and dietary supplements in health and disease. Journal of Food and Drug Analysis, 20(SUPPL.1), 226–230.
38. Shen, Q., Zhang, C., Jia, W., Qin, X., Cui, Z., & Mo, H. (2019). Co-production of chondroitin sulfate and peptide from lique fied chicken sternal cartilage by hot-pressure. Carbohydrate Polymers, 222(April), 115015. https://doi.org/10.1016/j.carbpol.2019.115015
39. Soua, L., Koubaa, M., Barba, F. J., Fakhfakh, J., Ghamgui, H. K., & Chaabouni, S. E. (2020). Water-Soluble Polysaccharides from Ephedra alata Stems: Structural characterization, functional properties, and antioxidant activity. Molecules, 25(9), 1–18. https://doi.org/10.3390/molecules25092210
40. Souissi, N., Boughriba, S., Abdelhedi, O., Hamdi, M., Jridi, M., Li, S., & Nasri, M. (2019a). Extraction, structural characterization, and thermal and biomedical properties of sulfated polysaccharides from razor clam Solen marginatus. RSC Advances, 9(20), 11538–11551. https://doi.org/10.1039/C9RA00959K
41. Trigui, I., Yaich, H., Sila, A., Cheikh-Rouhou, S., Bougatef, A., Blecker, C., Attia, H., & Ayadi, M. A. (2018). Physicochemical properties of water-soluble polysaccharides from black cumin seeds. International Journal of Biological Macromolecules, 117, 937–946. https://doi.org/10.1016/j.ijbiomac.2018.05.202
42. Wang, Y., Ghosh, S., & Nickerson, M. T. (2019). Effect of pH on the formation of electrostatic complexes between lentil protein isolate and a range of anionic polysaccharides, and their resulting emulsifying properties. Food Chemistry, 298(June), 125023. https://doi.org/10.1016/j.foodchem.2019.125023
43. Xie, J. H., Tang, W., Jin, M. L., Li, J. E., & Xie, M. Y. (2016). Recent advances in bioactive polysaccharides from Lycium barbarum , Zizyphus jujuba Mill, Plantago spp., and Morus spp.: Structures and functionalities. Food Hydrocolloids, 60(235), 148–160. https://doi.org/10.1016/j.foodhyd.2016.03.030
44. Xu, S. Y., Huang, X., & Cheong, K. L. (2017a). Recent advances in marine algae polysaccharides: Isolation, structure, and activities. Marine Drugs, 15(12), 1–16. https://doi.org/10.3390/md15120388
45. Yang, L., & Zhang, L. (2009). Chemical structural and chain conformational characterization of some bioactive polysaccharides isolated from natural sources. Carbohydrate Polymers, 76(3), 349–361. https://doi.org/10.1016/j.carbpol.2008.12.015
46. Ye, Q., Georges, N., & Selomulya, C. (2018). Microencapsulation of active ingredients in functional foods: From research stage to commercial food products. Trends in Food Science and Technology, 78(January), 167–179. https://doi.org/10.1016/j.tifs.2018.05.025
47. Yu, Y., Shen, M., Song, Q., & Xie, J. (2018). Biological activities and pharmaceutical applications of polysaccharide from natural resources: A review. Carbohydrate Polymers, 183(235), 91–101. https://doi.org/10.1016/j.carbpol.2017.12.009
48. Zamora-Sillero, J., Gharsallaoui, A., & Prentice, C. (2018). Peptides from Fish By-product Protein Hydrolysates and Its Functional Properties: an Overview. Marine Biotechnology, 20(2), 118–130. https://doi.org/10.1007/s10126-018-9799-3
49. Zhu, D. Y., Ma, Y. L., Wang, C. H., Wang, H., Ren, Y. F., Zhang, J. G., Thakur, K., & Wei, Z. J. (2017). Insights into physicochemical and functional properties of polysaccharides sequentially extracted from onion (Allium cepa). International Journal of Biological Macromolecules, 105, 1192–1201. https://doi.org/10.1016/j.ijbiomac.2017.07.164