Abstract
This study aimed to investigate the individual and synergistic antioxidant effects of dipeptides in vitro. Based on a study of amino acid antioxidant activities, 36 dipeptides were chosen. All their individual antioxidant effects were tested in in vitro antioxidant evaluation systems, including the DPPH, ABTS, ORAC, and FRAP assays. Based on the results of individual antioxidant effects of these dipeptides, some of them were chosen to study their synergistic antioxidant effects in the four antioxidant assays. The results of the DPPH assay showed that dipeptides containing Cys exhibited radical scavenging activity. The synergistic antioxidant assay indicated that LC and CH, LC and CN, LC and CE, CH and CN, CH and CE, and CN and CE have significant synergistic effects (p < 0.01) in the DPPH assay. Moreover, the results of the ABTS assay showed that 17 dipeptides have ABTS radical scavenging activities and that all of them contain Cys, Trp, and Tyr residues. Their synergistic antioxidant assay showed that CR and CH, YR and YK, YR and YN, and WK and IW showed highly significant synergistic antioxidant effects (p < 0.01). The results of the ORAC assay showed that 22 dipeptides possessed ORAC activities and that all of them have one or two Cys, His, Met, Trp, and Tyr residues. The synergistic effects of CH and HL, WK and WR, YK and WR, LC and FM, FM and CH, and WK and IW were significant (p < 0.01). Finally, in the FRAP assay, six dipeptides showed ferric reducing antioxidant activities and all of them contained the Cys residue. The synergistic antioxidant effects of HV and CN, CK and CN, CH and CN, and CE and CN were also significant (p < 0.01). These findings indicate that the individual and synergistic antioxidant effects of dipeptides are related to their constituent amino acids. These results may help study other individual and synergistic antioxidant effects of dipeptides.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
Abbreviations
- YN:
-
Tyr-Asn
- NY:
-
Asn-Tyr
- DY:
-
Asp-Tyr
- DN:
-
Asp-Asn
- YS:
-
Tyr-Ser
- SY:
-
Ser-Tyr
- DS:
-
Asp-Ser
- DG:
-
Asp-Gly
- FM:
-
Phe-Met
- DT:
-
Asp-Thr
- DE:
-
Asp-Glu
- CR:
-
Cys-Arg
- LC:
-
Leu-Cys
- LR:
-
Leu-Arg
- CK:
-
Cys-Lys
- WK:
-
Trp-Lys
- CH:
-
Cys-His
- CN:
-
Cys-Asn
- FI:
-
Phe-Ile
- WR:
-
Trp-Arg
- IW:
-
Ile-Trp
- YR:
-
Tyr-Arg
- YK:
-
Tyr-Lys
- RK:
-
Arg-Lys
- CE:
-
Cys-Glu
- SA:
-
Ser-Ala
- QE:
-
Gln-Glu
- PE:
-
Pro-Glu
- SE:
-
Ser-Glu
- PT:
-
Pro-Thr
- HK:
-
His-Lys
- VR:
-
Val-Arg
- HV:
-
His-Val
- HR:
-
His-Arg
- HY:
-
His-Tyr
- HL:
-
His-Leu
- ORAC:
-
Oxygen radical absorbance capacity
- PBS:
-
Phosphate buffer saline
- DPPH:
-
2,2-Diphenyl-1-picrylhydrazyl
- ABTS:
-
2,2′-Azino-bis(3-ethylbenzothiazoline-6-sulphonic acid) diammonium salt
- FRAP:
-
Ferric reducing antioxidant power
References
Aliaga C, Lissi EA (2000) Reactions of the radical cation derived from 2,2′-azinobis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS(.+)) with amino acids. Kinetics and mechanism. Can J Chem 78:1052–1059. https://doi.org/10.1139/cjc-78-8-1052
Ashok BT, Ahmad J, Ali R (1998) Immunochemical detection of oxidative DNA damage in cancer and aging using anti-reactive oxygen species modified DNA monoclonal antibody. Int J Biochem Cell Biol 30:1367–1377. https://doi.org/10.1016/s1357-2725(98)00100-9
Becker EM, Ntouma G, Skibsted LH (2007) Synergism and antagonism between quercetin and other chain-breaking antioxidants in lipid systems of increasing structural organisation. Food Chem 103:1288–1296. https://doi.org/10.1016/j.foodchem.2006.10.034
Benzie IF, Strain JJ (1996) The ferric reducing ability of plasma (FRAP) as a measure of “antioxidant power”: the FRAP assay. Anal Biochem 239:70–76. https://doi.org/10.1006/abio.1996.0292
Bougatef A, Nedjar-Arroume N, Manni L, Ravallec R, Barkia A, Guillochon D, Nasri M (2010) Purification and identification of novel antioxidant peptides from enzymatic hydrolysates of sardinelle (Sardinella aurita) by-products proteins. Food Chem 118:559–565. https://doi.org/10.1016/j.foodchem.2009.05.021
Dantzig AH, Bergin L (1990) Uptake of the cephalosporin, cephalexin, by a dipeptide transport carrier in the human intestinal cell line, Caco-2. Biochim Biophys Acta 1027:211–217. https://doi.org/10.1016/0005-2736(90)90309-c
Dudonne S, Vitrac X, Coutiere P, Woillez M, Merillon J-M (2009) Comparative study of antioxidant properties and total phenolic content of 30 plant extracts of industrial interest using DPPH, ABTS, FRAP, SOD, and ORAC assays. J Agric Food Chem 57:1768–1774. https://doi.org/10.1021/jf803011r
Farvin KHS, Andersen LL, Otte J, Nielsen HH, Jessen F, Jacobsen C (2016) Antioxidant activity of cod (Gadus morhua) protein hydrolysates: fractionation and characterisation of peptide fractions. Food Chem 204:409–419. https://doi.org/10.1016/j.foodchem.2016.02.145
Garrett AR, Weagel EG, Martinez AD, Heaton M, Robison RA, O’Neill KL (2014) A novel method for predicting antioxidant activity based on amino acid structure. Food Chem 158:490–496. https://doi.org/10.1016/j.foodchem.2014.02.102
Guo H, Kouzuma Y, Yonekura M (2009) Structures and properties of antioxidative peptides derived from royal jelly protein. Food Chem 113:238–245. https://doi.org/10.1016/j.foodchem.2008.06.081
Han C-H, Lin Y-S, Lin S-Y, Hou W-C (2014) Antioxidant and antiglycation activities of the synthesised dipeptide, Asn-Trp, derived from computer-aided simulation of yam dioscorin hydrolysis and its analogue, Gln-Trp. Food Chem 147:195–202. https://doi.org/10.1016/j.foodchem.2013.09.109
Hatano B, Kubo D, Tagaya H (2006) Study on the reactivity of diarylmethane derivatives in supercritical alcohols media: reduction of diarylmethanols and diaryl ketones to diarylmethanes using supercritical 2-propanol. Chem Pharm Bull 54:1304–1307. https://doi.org/10.1248/cpb.54.1304
Huang DJ, Ou BX, Prior RL (2005) The chemistry behind antioxidant capacity assays. J Agric Food Chem 53:1841–1856. https://doi.org/10.1021/jf030723c
Kumara HK, Gowda DC (2017) Synthesis and SAR studies of bisthiourea derivatives of dipeptides Lys/lys-Asp, Lys/lys-Trp conjugated benzo[d]isoxazole as promising antioxidants. Int J Pept Res Ther 23:259–267. https://doi.org/10.1007/s10989-016-9557-1
Liu J, Chen Z, He J, Zhang Y, Zhang T, Jiang Y (2014) Anti-oxidative and anti-apoptosis effects of egg white peptide, Trp-Asn-Trp-Ala-Asp, against H2O2-induced oxidative stress in human embryonic kidney 293 cells. Food Funct 5:3179–3188. https://doi.org/10.1039/c4fo00665h
Liu JB, Jin Y, Lin SY, Jones GS, Chen F (2015) Purification and identification of novel antioxidant peptides from egg white protein and their antioxidant activities. Food Chem 175:258–266. https://doi.org/10.1016/j.foodchem.2014.11.142
Mirzaei H, Regnier F (2008) Protein: protein aggregation induced by protein oxidation. J Chromatogr B 873:8–14. https://doi.org/10.1016/j.jchromb.2008.04.025
Newey H, Smyth DH (1960) Intracellular hydrolysis of dipeptides during intestinal absorption. J Physiol 152:367–380. https://doi.org/10.1113/jphysiol.1960.sp006493
Ou BX, Huang DJ, Hampschl-Woodill M, Flanagan JA (2003) When east meets west: the relationship between yin-yang and antioxidation-oxidation. FASEB J 17:127–129. https://doi.org/10.1096/fj.02-0527hyp
Puchalska P, Luisa Marina M, Concepcion Garcia M (2014) Isolation and identification of antioxidant peptides from commercial soybean-based infant formulas. Food Chem 148:147–154. https://doi.org/10.1016/j.foodchem.2013.10.030
Re R, Pellegrini N, Proteggente A, Pannala A, Yang M, Rice-Evans C (1999) Antioxidant activity applying an improved ABTS radical cation decolorization assay. Free Radical Biol Med 26:1231–1237. https://doi.org/10.1016/s0891-5849(98)00315-3
Sanchez-Moreno C (2002) Review: methods used to evaluate the free radical scavenging activity in foods and biological systems. Food Sci Technol Int 8:121–137. https://doi.org/10.1106/108201302026770
Seo H-S, Kwak S-Y, Lee Y-S (2010) Antioxidative activities of histidine containing caffeic acid-dipeptides. Bioorg Med Chem Lett 20:4266–4272. https://doi.org/10.1016/j.bmcl.2010.04.135
Thaipong K, Boonprakob U, Crosby K, Cisneros-Zevallos L, Byrne DH (2006) Comparison of ABTS, DPPH, FRAP, and ORAC assays for estimating antioxidant activity from guava fruit extracts. J Food Compos Anal 19:669–675. https://doi.org/10.1016/j.jfca.2006.01.003
Wang C, Wang B, Li B (2016) Bioavailability of peptides from casein hydrolysate in vitro: amino acid compositions of peptides affect the antioxidant efficacy and resistance to intestinal peptidases. Food Res Int 81:188–196. https://doi.org/10.1016/j.foodres.2015.12.013
Wiriyaphan C, Chitsomboon B, Roytrakul S, Yongsawadigul J (2013) Isolation and identification of antioxidative peptides from hydrolysate of threadfin bream surimi processing byproduct. J Funct Foods 5:1654–1664. https://doi.org/10.1016/j.jff.2013.07.009
Yan Q-J, Huang L-H, Sun Q, Jiang Z-Q, Wu X (2015) Isolation, identification and synthesis of four novel antioxidant peptides from rice residue protein hydrolyzed by multiple proteases. Food Chem 179:290–295. https://doi.org/10.1016/j.foodchem.2015.01.137
Zhang D, Wang Y, Xu M, Ding L, Zhang T, Liu J (2017) Antioxidant synergetic effect between the peptides derived from the egg white pentapeptide Trp-Asn-Trp-Ala-Asp international. J Pept Res Ther 8:1–10
Zheng L, Lin L, Su G, Zhao Q, Zhao M (2015) Pitfalls of using 1,1-diphenyl-2-picrylhydrazyl (DPPH) assay to assess the radical scavenging activity of peptides: its susceptibility to interference and low reactivity towards peptides. Food Res Int 76:359–365. https://doi.org/10.1016/j.foodres.2015.06.045
Zheng L, Zhao MM, Xiao CQ, Zhao QZ, Su GW (2016) Practical problems when using ABTS assay to assess the radical-scavenging activity of peptides: importance of controlling reaction pH and time. Food Chem 192:288–294. https://doi.org/10.1016/j.foodchem.2015.07.015
Zou TB, He TP, Li HB, Tang HW, Xia EQ (2016) The structure-activity relationship of the antioxidant peptides from natural proteins. Molecules 21:14. https://doi.org/10.3390/molecules21010072
Acknowledgements
The authors acknowledge the financial support provided by the National Natural Science foundation of China (31601486, 31771985), Fundamental Research Funds for the Central Universities (451170301197), Jilin Key Laboratory of Nutrition and Functional Food (20160622030JC), and the Program of JLU Science and Technology Innovative Research Team (JLUSTIRT).
Author information
Authors and Affiliations
Corresponding author
Ethics declarations
Conflict of interest
The authors declare no competing financial interests.
Rights and permissions
About this article
Cite this article
Du, Z., Liu, J., Zhang, D. et al. Individual and Synergistic Antioxidant Effects of Dipeptides in In Vitro Antioxidant Evaluation Systems. Int J Pept Res Ther 25, 391–399 (2019). https://doi.org/10.1007/s10989-018-9684-y
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s10989-018-9684-y