Plant Foods for Human Nutrition

, Volume 71, Issue 4, pp 402–409 | Cite as

Antioxidant and Anticancer Activities of Walnut (Juglans regia L.) Protein Hydrolysates Using Different Proteases

  • Raheleh Jahanbani
  • S. Mahmood Ghaffari
  • Maryam Salami
  • Kourosh Vahdati
  • Houri Sepehri
  • Nazanin Namazi Sarvestani
  • Nader Sheibani
  • Ali Akbar Moosavi-MovahediEmail author
Original Paper


Walnut (Juglans regia L.) contains approximately 20–25 % protein with abundant essential amino acids. The enzymatic hydrolysate of Persian walnut (Chandler) seed proteins was prepared by incubation with three different proteases, including pancreatic chymotrypsin and trypsin, and a microbial enzyme proteinase K. The hydrolysates were found to possess excellent antioxidant capacities. The peptide fractions scavenged the 2, 2′-anizo-bis-(3-ethylbenzthiazoline-6-sulfonic acid) (ABTS) free radicals and inhibited the activity of reactive oxygen species. Walnut protein hydrolysates were also tested, for the first time, against the viability of human breast (MDA-MB231) and colon (HT-29) cancer cell lines. MTT, [3-(4, 5dimethylthiazolyl)-2,5-diphenyl-tetrazolium bromide], assay was used to assess in vitro cancer cell viability upon treatment with the peptide fractions. The peptide fractions showed cell growth inhibition of 63 ± 1.73 % for breast cancer and 51 ± 1.45 % for colon cancer cells. Thus, a direct correlation between antioxidant and anticancer activities of walnut peptide fractions exists and supports their potential therapeutic benefit.


Walnut proteins Bioactive peptides Antioxidant Anticancer Protease 



2, 2′-anizo-bis-(3-ethylbenzthiazoline-6-sulfonic acid)


Angiotensin converting enzyme




Human umbilical vein endothelial cells


3-(4, 5dimethylthiazolyl)-2,5-diphenyl-tetrazolium bromide




Reactive oxygen species


Sodium dodecyl sulfate



The support of University of Tehran, International Scientific Studies & Collaboration (CISSC)-Ministry of Science, Research and Technology in Iran, Center of Excellence in Biothermodynamics (CEBiotherm), Center of Excellence for Walnut Improvement and Technology of Iran, Iran National Science Foundation (INSF) and Iran National Elites Foundation (INEF) and UNESCO Chair on Interdisciplinary Research in Diabetes, Iran Society of Biophysical Chemistry is gratefully acknowledged. The authors also acknowledge the Headquarter of Science and Technology Development in Medicinal Plants of Iran’s Vice-President for Science and Technology Affairs.

Compliance with Ethical Standards

Conflict of Interest

The authors declare that there are no conflicts of interest.

Human and Animal Rights

This article does not contain any studies with human or animal subjects.


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Copyright information

© Springer Science+Business Media New York 2016

Authors and Affiliations

  • Raheleh Jahanbani
    • 1
  • S. Mahmood Ghaffari
    • 1
  • Maryam Salami
    • 2
  • Kourosh Vahdati
    • 3
  • Houri Sepehri
    • 4
  • Nazanin Namazi Sarvestani
    • 5
  • Nader Sheibani
    • 4
  • Ali Akbar Moosavi-Movahedi
    • 1
    • 6
    Email author
  1. 1.Institute of Biochemistry and BiophysicsUniversity of TehranTehranIran
  2. 2.Department of Food Science and Engineering, University College of Agriculture & Natural ResourcesUniversity of TehranKarajIran
  3. 3.Department of Horticulture, College of AburaihanUniversity of TehranTehranIran
  4. 4.Animal Biology Department, School of Biology, College of SciencesUniversity of TehranTehranIran
  5. 5.Departments of Ophthalmology and Visual Sciences, and Biomedical EngineeringUniversity of Wisconsin School of Medicine and Public HealthMadisonUSA
  6. 6.Center of Excellence in BiothermodynamicsUniversity of TehranTehranIran

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