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Biological Potential of Flaxseed Protein Hydrolysates Obtained by Different Proteases

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Abstract

Flaxseed meal, a byproduct of flaxseed oil extraction, was treated as low-value agrowaste for a long time despite its high protein content. Flaxseed meal has recently garnered increasing interest as a source of proteins and other bioactive compounds with positive impacts on human health. The aim of this study was to investigate the in vitro biological potential of flaxseed protein hydrolysates (FPH). Three FPHs were prepared using three hydrolytic enzymes: Alcalase, Neutrase and Protamex. The molecular weight profile of peptides contained in the hydrolysates was determined by size exclusion chromatography (SEC). The oxygen radical absorbance capacity (ORAC) assay was used to determine the peptide antioxidant capacity, while proliferative effects were studied in two cell lines: HeLa and HaCaT. The latter was also used to determine the protective effect of the FPH during induced oxidative stress. Alcalase showed the highest proteolytic activity, while the produced flaxseed protein hydrolysate (FPH-A) exhibited the strongest antioxidant potential. FPH-A had cytotoxic effects at 10 mg/mL in HeLa cells, but it stimulated HaCaT cell growth. Moreover, a mild protective effect of FPH-A was detected in HaCaT cells after induction of oxidative stress. Flaxseed protein hydrolysates obtained by Neutrase (FPH-N) and Protamex (FPH-P) have less pronounced or no potential at all, with respect to their antioxidative or antiproliferative activity. Therefore, to increase value-added utilization of flaxseed meal we suggest further research on hydrolysate obtained by Alcalase.

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Abbreviations

DCF-DA:

Dichlorohydrofluorescein-diacetate

DME:

Dulbecco’s modified Eagle’s medium

FBS:

Fetal bovine serum

FPH:

Flaxseed protein hydrolysate

FPH-A:

Flaxseed protein hydrolysate obtained by Alcalase

FPH-N:

Flaxseed protein hydrolysate obtained by Neutrase

FPH-P:

Flaxseed protein hydrolysate obtained by Protamex

FPI:

Flaxseed protein isolate

MTS:

3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium

ORAC:

Oxygen radical absorbance capacity

ROS:

Reactive oxygen species

SEC:

Size exclusion chromatography

References

  1. Oomah BD (2020) Flaxseed by-products. In: Campos-Vega R, Oomah BD, Vergara-Castañeda HA (eds) Food wastes and by-products: nutraceutical and health potential, 1st edn. Wiley, New York, pp 267–289

    Chapter  Google Scholar 

  2. Marambe PWMLHK, Wanasundara JPD (2017) Protein from flaxseed (Linum usitatissimun L). In: Nadathur SR, Wanasundara JPD, Scanlin L (eds) Sustainable protein sources, 1st edn. Academic Press, San Diego, pp 133–144

    Chapter  Google Scholar 

  3. Wu S, Wang X, Qi W, Guo Q (2019) Bioactive protein/peptides of flaxseed: a review. Trends Food Sci Tech 92:184–193. https://doi.org/10.1016/j.tifs.2019.08.017

    Article  CAS  Google Scholar 

  4. Marambe PWMLH, Shand PJ, Wanasundara JPD (2008) An in vitro investigation of selected biological activities of hydrolysed flaxseed (Linum usitatissimum L.) proteins. J Am Oil Chem Soc 85:1155–1164. https://doi.org/10.1007/s11746-008-1293-z

    Article  CAS  Google Scholar 

  5. Udenigwe CC, Lu YL, Han CH, Hou WC, Aluko RE (2009) Flaxseed protein-derived peptide fractions: antioxidant properties and inhibition of lipopolysaccharide-induced nitric oxide production in murine macrophages. Food Chem 116:277–284. https://doi.org/10.1016/j.foodchem.2009.02.046

    Article  CAS  Google Scholar 

  6. Udenigwe CC, Aluko RE (2012) Food protein-derived bioactive peptides: production, processing, and potential health benefits. J Food Sci 71:11–24. https://doi.org/10.1111/j.1750-3841.2011.02455.x

    Article  CAS  Google Scholar 

  7. Udenigwe CC, Aluko RE (2012) Multifunctional cationic peptide fractions from flaxseed protein hydrolysates. Plant Foods Hum Nutr 67:1–9. https://doi.org/10.1007/s11130-012-0275-3

  8. Nwachukwu ID, Aluko RE (2018) Antioxidant properties of flaxseed protein hydrolyzates: influence of hydrolytic enzyme concentration and peptide size. J Am Oil Chem Soc 95:1105–1118. https://doi.org/10.1002/aocs.12042

    Article  CAS  Google Scholar 

  9. Karamać M, Kulczyk A, Sulewska K (2014) Antioxidant activity of hydrolysates prepared from flaxseed cake proteins using pancreatin. Pol J Food Nutr Sci 64:227–233. https://doi.org/10.2478/pjfns-2013-0023

    Article  CAS  Google Scholar 

  10. Karamać M, Kosinska-Cagnazzo A, Kulczyk A (2016) Use of different proteases to obtain flaxseed protein hydrolysates with antioxidant activity. Int J Mol Sci 17:1027–1040. https://doi.org/10.3390/ijms17071027

    Article  CAS  PubMed Central  Google Scholar 

  11. Silva FGD, Hernández-Ledesma B, Amigo L, Netto FM, Miralles B (2016) Identification of peptides released from flaxseed (Linum usitatissimum) protein by Alcalase® hydrolysis: antioxidant activity. LWT – Food Sci Technol 76:140–146. https://doi.org/10.1016/j.lwt.2016.10.049

    Article  CAS  Google Scholar 

  12. Sarmadi BH, Ismail A (2010) Antioxidative peptides from food proteins: a review. Peptides 31:1949–1956. https://doi.org/10.1016/j.peptides.2010.06.020

    Article  CAS  PubMed  Google Scholar 

  13. Tang CH, Wang XS, Yang XQ (2009) Enzymatic hydrolysis of hemp (Cannabis sativa L.) protein isolate by various proteases and antioxidant properties of the resulting hydrolysates. Food Chem 114:1484–1490. https://doi.org/10.1016/j.foodchem.2008.11.049

    Article  CAS  Google Scholar 

  14. Zhang Q, Tong X, Qi B, Wang Z, Li Y, Sui J, Jiang L (2018) Changes in antioxidant activity of Alcalase-hydrolyzed soybean hydrolysate under simulated gastrointestinal digestion and transepithelial transport. J Funct Foods 42:298–305. https://doi.org/10.1016/j.jff.2018.01.017

    Article  CAS  Google Scholar 

  15. Onuh JO, Girgih AT, Aluko RE, Aliani M (2014) In vitro antioxidant properties of chicken skin enzymatic protein hydrolysates and membrane fractions. Food Chem 150:366–373. https://doi.org/10.1016/j.foodchem.2013.10.107

    Article  CAS  PubMed  Google Scholar 

  16. He R, Alashi A, Malomo SA, Girgih AT, Chao D, Ju X, Aluko RE (2013) Antihypertensive and free radical scavenging properties of enzymatic rapeseed protein hydrolysates. Food Chem 141:153–159. https://doi.org/10.1016/j.foodchem.2013.02.087

    Article  CAS  PubMed  Google Scholar 

  17. González Montoya M, Ramón-Gallegos E, Robles-Ramírez MC, Mora-Escobedo R (2016) Evaluation of the antioxidant and antiproliferative effects of three peptide fractions of germinated soybeans on breast and cervical cancer cell lines. Plant Foods Hum Nutr 71:368–374. https://doi.org/10.1007/s11130-016-0568-z

  18. Girón-Cale J, Alaiz M, Vioque J (2010) Effect of chickpea protein hydrolysates on cell proliferation and in vitro bioavailability. Food Res Int 43:1365–1370. https://doi.org/10.1016/j.foodres.2010.03.020

    Article  CAS  Google Scholar 

  19. Jahanbani R, Ghaffari SM, Salami M, Vahdan K, Sephri H, Sarvestani NN, Sheibani N, Moosavi-Movahedi AA (2016) Antioxidant and anticancer activities of walnut (Juglans regia L.) protein hydrolysates using different proteases. Plant Foods Hum Nutr 71:402–409. https://doi.org/10.1007/s11130-016-0576-z

  20. Logarušić M, Slivac I, Radošević K, Bagović M, Radojčić Redovniković I, Gaurina Srček V (2019) Hempseed protein hydrolysates’ effects on the proliferation and induced oxidative stress in normal and cancer cell lines. Mol Biol Rep 46:6079–6085. https://doi.org/10.1007/s11033-019-05043-8

    Article  CAS  PubMed  Google Scholar 

  21. Lu RR, Qian P, Sun Z, Zhou XH, Chen TP, He JF, Zhang H, Wu J (2010) Hempseed protein derived antioxidative peptides: purification, identification and protection from hydrogen peroxide-induced apoptosis in PC12 cells. Food Chem 123:1210–1218. https://doi.org/10.1016/j.foodchem.2010.05.089

    Article  CAS  Google Scholar 

  22. Silva FGD, O'Callagahan Y, O'Brien M, Netto FM (2013) Antioxidant capacity of flaxseed products: the effect of in vitro digestion. Plant Foods Hum Nutr 68:24–30. https://doi.org/10.1007/s11130-012-0329-6

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Acknowledgments

This work was funded by the Croatian Science Foundation HRZZ (Grant No. IP-2016-06-3848).

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Authors and Affiliations

  1. Faculty of Food Technology and Biotechnology, Laboratory for Cell Technology and Biotransformation, University of Zagreb, Pierottijeva 6, 10000, Zagreb, Croatia

    Marijan Logarušić, Kristina Radošević, Manuela Panić, Igor Slivac & Višnja Gaurina Srček

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  1. Marijan Logarušić
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  2. Kristina Radošević
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  3. Ana Bis
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  4. Manuela Panić
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  5. Igor Slivac
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  6. Višnja Gaurina Srček
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Correspondence to Višnja Gaurina Srček.

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Logarušić, M., Radošević, K., Bis, A. et al. Biological Potential of Flaxseed Protein Hydrolysates Obtained by Different Proteases. Plant Foods Hum Nutr 75, 518–524 (2020). https://doi.org/10.1007/s11130-020-00841-z

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