Skip to main content

Advertisement

SpringerLink
Log in

Identification of Two Novel Antioxidant Peptides from Camel Milk Using Digestive Proteases: Impact on Expression Gene of Superoxide Dismutase (SOD) in Hepatocellular Carcinoma Cell Line

  • Published:
International Journal of Peptide Research and Therapeutics Aims and scope Submit manuscript

Abstract

Camel milk has high levels of antioxidant peptides; therefore has a significant role in nutrition and health. There is a wide range of diseases associated with oxidative stress in the body, thereby preventing the production of free radicals followed the occurrence of such reactions have an important role in human health. Some peptides derived from natural sources as natural antioxidants have a significant role in the inhibition of free radicals. The aim of this study was the identification and purification of camel milk proteins and evaluation of their antioxidant properties. For this purpose, camel milk proteins were hydrolyzed and examined the antioxidant properties of resulting peptides. Ultrafiltration and reverse-phase HPLC techniques were used for fractionation of the hydrolysate. Antioxidant activity of peptides were evaluated using different methods such as 2,2′-diphenyl-1-picrylhydrazyl (DPPH·), 2,2′-azinobis(3-ethylbenzothiazo-line-6-sulfonic acid) diammonium salt (ABTS+), superoxide (O2·−), hydroxyl (OH·−) and polyunsaturated fatty acid peroxidation assays. The real-time PCR was performed to evaluate changes in expression of superoxide dismutase gene (SOD) as an antioxidant enzyme in HepG2 cells treated with different concentrations of peptide. Active peaks were analyzed using tandem mass spectrometry and thus identified two peptides NEDNHPGALGEPV (NV-13) and KVLPVPQQMVPYPRQ (KQ-15) with 1348.38 and 1780.15 dalton molecular weight, respectively. Both peptides showed antioxidant effects but KQ-15 peptide showed stronger effects than other. Molecular analysis showed that KQ-15 peptide was also able to increase the expression of SOD gene. The results show that the NV-13 and KQ-15 peptides have a high antioxidant capacity and they can be used to deal with diseases associated with oxidative stress.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6
Fig. 7

Similar content being viewed by others

References

  • Aloğlu HŞ, Öner Z (2011) Determination of antioxidant activity of bioactive peptide fractions obtained from yogurt. J Dairy Sci 94:5305–5314

    Article  PubMed  Google Scholar 

  • Asoodeh A, Naderi-Manesh H, Mirshahi M, Ranjbar B (2004) Purification and characterization of an antibacterial, antifungal and non hemolytic peptide from Rana Ridibunda. J Sci Islam Repub Iran 15:303–309

    Google Scholar 

  • Aydin S, Aydin S, Ozkan Y, Kumru S (2006) Ghrelin is present in human colostrum, transitional and mature milk. Peptides 27:878–882

    Article  CAS  PubMed  Google Scholar 

  • Binsan W, Benjakul S, Visessanguan W, Roytrakul S, Tanaka M, Kishimura H (2008) Antioxidative activity of Mungoong, an extract paste, from the cephalothorax of white shrimp (Litopenaeus vannamei). Food Chem 106:185–193

    Article  CAS  Google Scholar 

  • Byun HG, Lee JK, Park HG, Jeon JK, Kim SK (2009) Antioxidant peptides isolated from the marine rotifer, Brachionus rotundiformis. Process Biochem 44:842–846

    Article  CAS  Google Scholar 

  • Cacciuttolo MA, Trinh L, Lumpkin JA, Rao G (1993) Hyperoxia induces DNA damage in mammalian cells. Free Radic Biol Med 14:267–276

    Article  CAS  PubMed  Google Scholar 

  • Castro MM, Fandiño RLA, Sanchez MIR, Gonzalez MMR, De Artiñano AA (2012) Bioactive peptides derived from the proteins of egg white by means of enzymatic hydrolysis. Google Patents. US8227207 B2

  • De Gobba C, Espejo-Carpio FJ, Skibsted LH, Otte J (2014) Antioxidant peptides from goat milk protein fractions hydrolysed by two commercial proteases. Int Dairy J 39:28–40

    Article  Google Scholar 

  • El Agamy ESI, Ruppanner R, Ismail A, Champagne CP, Assaf R (1992) Antibacterial and antiviral activity of camel milk protective proteins. J Dairy Res 59:169–175

    Article  PubMed  Google Scholar 

  • Ganapathy E, Su F, Meriwether D, Devarajan A, Grijalva V, Gao F, Chattopadhyay A, Anantharamaiah G, Navab M, Fogelman AM (2012) D-4F, an apoA-I mimetic peptide, inhibits proliferation and tumorigenicity of epithelial ovarian cancer cells by upregulating the antioxidant enzyme MnSOD. Int J Cancer 130:1071–1081

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • 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

    Article  CAS  Google Scholar 

  • Hernández-Ledesma B, Quirós A, Amigo L, Recio I (2007) Identification of bioactive peptides after digestion of human milk and infant formula with pepsin and pancreatin. Int Dairy J 17:42–49

    Article  Google Scholar 

  • Je JY, Park PJ, Kim SK (2005) Antioxidant activity of a peptide isolated from Alaska pollack (Theragra chalcogramma) frame protein hydrolysate. Food Res Int 38:45–50

    Article  CAS  Google Scholar 

  • Jun SY, Park PJ, Jung WK, Kim SK (2004) Purification and characterization of an antioxidative peptide from enzymatic hydrolysate of yellowfin sole (Limanda aspera) frame protein. Eur Food Res Technol 219:20–26

    Article  CAS  Google Scholar 

  • Li YW, Li B (2013) Characterization of structure–antioxidant activity relationship of peptides in free radical systems using QSAR models: key sequence positions and their amino acid properties. J Theor Biol 318:29–43

    Article  CAS  PubMed  Google Scholar 

  • Li Q, Dong C, Deng A, Katsumata M, Nakadai A, Kawada T, Okada S, Clayberger C, Krensky AM (2005) Hemolysis of erythrocytes by granulysin-derived peptides but not by granulysin. Antimicrob Agents Chemother 49:388–397

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Li L, Ng T, Song M, Yuan F, Liu Z, Wang C, Jiang Y, Fu M, Liu F (2007) A polysaccharide–peptide complex from abalone mushroom (Pleurotus abalonus) fruiting bodies increases activities and gene expression of antioxidant enzymes and reduces lipid peroxidation in senescence-accelerated mice. Appl Microbiol Biotechnol 75:863–869

    Article  CAS  PubMed  Google Scholar 

  • Li Y, Jiang B, Zhang T, Mu W, Liu J (2008) Antioxidant and free radical-scavenging activities of chickpea protein hydrolysate (CPH). Food Chem 106:444–450

    Article  CAS  Google Scholar 

  • Li J, Li Q, Li J, Zhou B (2014) Peptides derived from Rhopilema esculentum hydrolysate exhibit angiotensin converting enzyme (ACE) inhibitory and antioxidant abilities. Molecules 19:13587–13602

    Article  PubMed  Google Scholar 

  • Liu Q, Kong B, Xiong YL, Xia X (2010) Antioxidant activity and functional properties of porcine plasma protein hydrolysate as influenced by the degree of hydrolysis. Food Chem 118:403–410

    Article  CAS  Google Scholar 

  • Mendis E, Rajapakse N, Kim SK (2005) Antioxidant properties of a radical-scavenging peptide purified from enzymatically prepared fish skin gelatin hydrolysate. J Agric Food Chem 53:581–587

    Article  CAS  PubMed  Google Scholar 

  • Peng X, Xiong YL, Kong B (2009) Antioxidant activity of peptide fractions from whey protein hydrolysates as measured by electron spin resonance. Food Chem 113:196–201

    Article  CAS  Google Scholar 

  • Pownall TL, Udenigwe CC, Aluko RE (2010) Amino acid composition and antioxidant properties of pea seed (Pisum sativum L.) enzymatic protein hydrolysate fractions. J Agric Food Chem 58:4712–4718

    Article  CAS  PubMed  Google Scholar 

  • Qian ZJ, Jung WK, Byun HG, Kim SK (2008) Protective effect of an antioxidative peptide purified from gastrointestinal digests of oyster, Crassostrea gigas against free radical induced DNA damage. Bioresour Technol 99:3365–3371

    Article  CAS  PubMed  Google Scholar 

  • Quirós A, del Mar Contreras M, Ramos M, Amigo L, Recio I (2009) Stability to gastrointestinal enzymes and structure–activity relationship of β-casein-peptides with antihypertensive properties. Peptides 30:1848–1853

    Article  PubMed  Google Scholar 

  • Rajapakse N, Mendis E, Byun HG, Kim SK (2005) Purification and in vitro antioxidative effects of giant squid muscle peptides on free radical-mediated oxidative systems. J Nutr Biochem 16:562–569

    Article  CAS  PubMed  Google Scholar 

  • Saiga A, Tanabe S, Nishimura T (2003) Antioxidant activity of peptides obtained from porcine myofibrillar proteins by protease treatment. J Agric Food Chem 51:3661–3667

    Article  CAS  PubMed  Google Scholar 

  • Salami M, Yousefi R, Ehsani MR, Razavi SH, Chobert JM, Haertlé T, Saboury AA, Atri MS, Niasari-Naslaji A, Ahmad F (2009) Enzymatic digestion and antioxidant activity of the native and molten globule states of camel α-lactalbumin: possible significance for use in infant formula. Int Dairy J 19:518–523

    Article  CAS  Google Scholar 

  • Sheu SS, Nauduri D, Anders M (2006) Targeting antioxidants to mitochondria: a new therapeutic direction. Biochim Biophys Acta (BBA) 1762:256–265

    Article  CAS  Google Scholar 

  • Sila A, Sayari N, Balti R, Martinez-Alvarez O, Nedjar-Arroume N, Moncef Bougatef A (2014) Biochemical and antioxidant properties of peptidic fraction of carotenoproteins generated from shrimp by-products by enzymatic hydrolysis. Food Chem 148:445–452

    Article  CAS  PubMed  Google Scholar 

  • Stadtman ER (1992) Protein oxidation and aging. Science 257:1220–1224

    Article  CAS  PubMed  Google Scholar 

  • Taheri A, Farvin KS, Jacobsen C, Baron CP (2014) Antioxidant activities and functional properties of protein and peptide fractions isolated from salted herring brine. Food Chem 142:318–326

    Article  CAS  PubMed  Google Scholar 

  • You L, Zhao M, Regenstein JM, Ren J (2011) In vitro antioxidant activity and in vivo anti-fatigue effect of loach (Misgurnus anguillicaudatus) peptides prepared by papain digestion. Food Chem 124:188–194

    Article  CAS  Google Scholar 

  • Zhang QX, Wu H, Ling YF, Lu RR (2013) Isolation and identification of antioxidant peptides derived from whey protein enzymatic hydrolysate by consecutive chromatography and Q-TOF MS. J Dairy Res 80:367–373

    Article  CAS  PubMed  Google Scholar 

Download references

Acknowledgments

This work was supported by Science and Research Branch, Islamic Azad University, Mashhad, Iran (Grant number: 75.304.92439) and Ferdowsi University of Mashhad, Mashhad, Iran and therefore is appreciated by the authors.

Author information

Authors and Affiliations

  1. Department of Biochemistry and Biophysics, Mashhad Branch, Islamic Azad University, Mashhad, Iran

    Masoud Homayouni-Tabrizi

  2. Department of Basic Science, Faculty of Veterinary Medicine, Ferdowsi University of Mashhad, Mashhad, Iran

    Hoda Shabestarin

  3. Department of Chemistry, Faculty of Science, Ferdowsi University of Mashhad, Mashhad, Iran

    Ahmad Asoodeh

  4. Department of Biology, Mashhad Branch, Islamic Azad University, Mashhad, Iran

    Mozhgan Soltani

Authors
  1. Masoud Homayouni-Tabrizi
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Hoda Shabestarin
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Ahmad Asoodeh
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Mozhgan Soltani
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Ahmad Asoodeh.

Ethics declarations

Conflict of Interest

Masoud Homayouni-Tabrizi, Ahmad Asoodeh, Mozhgan Soltani and Hoda Shabestarin declare that they have no conflict of interest.

Human and Animal Rights and Informed Consent

This article does not contain any studies with human or animal subjects performed by any of the authors.

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Homayouni-Tabrizi, M., Shabestarin, H., Asoodeh, A. et al. Identification of Two Novel Antioxidant Peptides from Camel Milk Using Digestive Proteases: Impact on Expression Gene of Superoxide Dismutase (SOD) in Hepatocellular Carcinoma Cell Line. Int J Pept Res Ther 22, 187–195 (2016). https://doi.org/10.1007/s10989-015-9497-1

Download citation

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s10989-015-9497-1

Keywords

Search

Navigation