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Hempseed protein hydrolysates’ effects on the proliferation and induced oxidative stress in normal and cancer cell lines

  • Marijan Logarušić
  • Igor Slivac
  • Kristina Radošević
  • Martina Bagović
  • Ivana Radojčić Redovniković
  • Višnja Gaurina SrčekEmail author
Original Article
  • 39 Downloads

Abstract

Food proteins from different sources can provide beneficial effects on human health by releasing the bioactive peptides that are integral part of their native structure. In this study, we tested the biological potential of hempseed protein hydrolysates (HPHs) obtained from hempseed cake protein isolate. The HPHs were prepared by enzyme hydrolysis using three different proteases of microbial origin: Alcalase®, Neutrase® and Protamex®. The antioxidant activity of the obtained hydrolysates was determined by oxygen radical absorbance capacity (ORAC) assay, while the proliferative effects on normal (HaCaT) and cancer (HeLa) cells were determined by the CellTiter 96® AQueous One Solution Reagent (MTS) assay. HPHs showed dose-dependent antiproliferative effects on HeLa cells and stimulatory effects on the proliferation of HaCaT cells. HPH obtained by Neutrase® (HPH-N) showed the highest antioxidant activity expressed as an ORAC value. The protective effect of HPH-N on H2O2-induced oxidative stress in normal and cancer cells was evaluated and 1 mg/mL of HPH-N significantly reduced the formation of intracellular reactive oxygen species (ROS) in both cell lines. The obtained results indicate the benefits of HPHs as potential natural antioxidants for the food industry and contribute to the growing trend of utilizing hempseed by-products.

Keywords

Antioxidant activity Antiproliferative effects Hempseed protein hydrolysates ORAC value 

Notes

Acknowledgements

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

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.

References

  1. 1.
    Kadam D, Lele SS (2018) Value addition of oilseed meal: a focus on bioactive peptides. Food Meas 12:449–458.  https://doi.org/10.1007/s11694-017-9658-3 CrossRefGoogle Scholar
  2. 2.
    Chalamaiah M, Yu W, Wu J (2018) Immunomodulatory and anticancer protein hydrolysates (peptides) from food proteins: a review. Food Chem 245:205–222.  https://doi.org/10.1016/j.foodchem.2017.10.087 CrossRefGoogle Scholar
  3. 3.
    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 CrossRefGoogle Scholar
  4. 4.
    Callaway JC (2004) Hempseed as a nutritional resource: an overview. Euphytica 140:65–72.  https://doi.org/10.1007/s10681-004-4811-6 CrossRefGoogle Scholar
  5. 5.
    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 CrossRefGoogle Scholar
  6. 6.
    Girgih AT, Udenigwe CC, Aluko RE (2011) In Vitro antioxidant properties of hemp seed (Cannabis sativa L.) protein hydrolysate fractions. J Am Oil Chem Soc 88:381–389.  https://doi.org/10.1007/s11746-010-1686-7 CrossRefGoogle Scholar
  7. 7.
    Wang XS, Tang CH, Chen L, Yang XQ (2009) Characterization and antioxidant properties of hemp protein hydrolysates obtained with Neutrase. Food Technol Biotechnol 47:428–434Google Scholar
  8. 8.
    Hong S, Sowndhararajan K, Joo T, Lim C, Cho H, Kim S, Kim GY, Jhoo JW (2015) Ethanol and supercritical fluid extracts of hemp seed (Cannabis sativa L.) increase gene expression of antioxidant enzymes in HepG2 cells. Asian Pac J Reprod 4:147–152.  https://doi.org/10.1016/s2305-0500(15)30012-9 CrossRefGoogle Scholar
  9. 9.
    Ninfali P, Mea G, Giorgini S, Rocchi M, Bacchiocca M (2005) Antioxidant capacity of vegetables, spices and dressings relevant to nutrition. Methods Enzymol 93:257–266.  https://doi.org/10.1079/bjn20041327 Google Scholar
  10. 10.
    Girgih AT, Udenigwe CC, Aluko RE (2013) Reverse-phase HPLC separation of hemp seed (Cannabis sativa L.) protein hydrolysate produced peptide fractions with enhanced antioxidant capacity. Plant Foods Hum Nutr 68:39–46.  https://doi.org/10.1007/s11130-013-0340-6 CrossRefGoogle Scholar
  11. 11.
    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 CrossRefGoogle Scholar
  12. 12.
    Kim S, Lee G (2009) Development of serum-free medium supplemented with hydrolysates for the production of therapeutic antibodies in CHO cell cultures using design of experiments. Appl Microbiol Biotechnol 83:639–648.  https://doi.org/10.1007/s00253-009-1903-1 CrossRefGoogle Scholar
  13. 13.
    Girón-Cale J, Vioque J, Pedroche J, Alaiz M, Yust MM, Megías C, Millán F (2008) Chickpea protein hydrolysate as a substitute for serum in cell culture. Cytotechnology 57:263–272.  https://doi.org/10.1007/s10616-008-9170-z CrossRefGoogle Scholar
  14. 14.
    Radošević K, Dukić B, Andlar M, Slivac I, Gaurina Srček V (2016) Adaptation and cultivation of permanent fish cell line CCO in serum-free medium and influence of protein hydrolysates on growth performance. Cytotechnology 68:115–121.  https://doi.org/10.1007/s10616-014-9760-x CrossRefGoogle Scholar
  15. 15.
    Mora-Escobedo R, Robles-Ramírez MC, Ramón-Gallegos E, Reza-Alemán R (2009) Effect of protein hydrolysates from germinated soybean on cancerous cells of the human cervix: an in vitro study. Plant Foods Hum Nutr 64:271–278.  https://doi.org/10.1007/s11130-009-0131-2 CrossRefGoogle Scholar
  16. 16.
    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 CrossRefGoogle Scholar
  17. 17.
    Wang L, Zhang J, Yuan Q, Xie H, Shi J, Ju X (2016) Separation and purification of an anti-tumor peptide from rapeseed (Brassica campestris L.) and the effect on cell apoptosis. Food Funct 7:2239–2248.  https://doi.org/10.1039/C6FO00042H CrossRefGoogle Scholar
  18. 18.
    Udenigwe CC, Aluko RE (2012) Food protein-derived bioactive peptides: production, processing, and potential health benefits. J Food Sci 71:R11–R24.  https://doi.org/10.1111/j.1750-3841.2011.02455.x CrossRefGoogle Scholar
  19. 19.
    Zhang QX, Ling YF, Sun Z, Zhang L, Yu HX, Kamau SM, Lu RR (2012) Protective effect of whey protein hydrolysates against hydrogen peroxide-induced oxidative stress on PC12 cells. Biotechnol Lett 34:2001–2006.  https://doi.org/10.1007/s10529-012-1017-1 CrossRefGoogle Scholar
  20. 20.
    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 CrossRefGoogle Scholar

Copyright information

© Springer Nature B.V. 2019

Authors and Affiliations

  1. 1.Laboratory for Cell Technology and Biotransformation, Faculty of Food Technology and BiotechnologyUniversity of ZagrebZagrebCroatia

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