Hostname: page-component-586b7cd67f-gb8f7 Total loading time: 0 Render date: 2024-11-22T18:23:41.420Z Has data issue: false hasContentIssue false

Isolation and identification of antioxidant peptides derived from whey protein enzymatic hydrolysate by consecutive chromatography and Q-TOF MS

Published online by Cambridge University Press:  22 July 2013

Qiu-Xiang Zhang
Affiliation:
State Key Laboratory of Food Science and Technology, School of Food Science and Technology, Jiangnan University, 1800 Lihu Avenue, Wuxi, Jiangsu 214122, China
Hui Wu
Affiliation:
State Key Laboratory of Food Science and Technology, School of Food Science and Technology, Jiangnan University, 1800 Lihu Avenue, Wuxi, Jiangsu 214122, China
Yu-Fang Ling
Affiliation:
State Key Laboratory of Food Science and Technology, School of Food Science and Technology, Jiangnan University, 1800 Lihu Avenue, Wuxi, Jiangsu 214122, China
Rong-Rong Lu*
Affiliation:
State Key Laboratory of Food Science and Technology, School of Food Science and Technology, Jiangnan University, 1800 Lihu Avenue, Wuxi, Jiangsu 214122, China
*
*For correspondence; e-mail: [email protected]

Abstract

To isolate and identify antioxidant peptides from enzymatically hydrolysed whey protein, whey protein isolate was hydrolysed by different protease (trypsin, pepsin, alcalase 2·4L, promatex, flavourzyme, protease N). The hydrolysate generated by alcalase 2·4L had the highest antioxidant activities on 1,1-diphenyl-2-picrylhydrazyl (DPPH) radicals, superoxide radicals and in a linoleic acid peroxidation system induced by Fe2+. The IC50 values of DPPH and superoxide radical scavenging activities of the hydrolysate decreased significantly (6·89 and 38·88%, respectively) after treatment with macroporous adsorption resin. Seven different peptides showing strong antioxidant activities were isolated from the hydrolysate using consecutive chromatographic methods including gel filtration chromatography and high-performance liquid chromatography. The molecular mass and amino acids sequences of the purified peptides were determined using a Quadrupole time-of-flight mass spectrometer (Q-TOF MS). One of the antioxidative peptides, Trp–Tyr–Ser–Leu, displayed the highest DPPH radical scavenging activity (IC50=273·63 μm) and superoxide radical scavenging activity (IC50=558·42 μm). These results suggest that hydrolysates from whey proteins are good potential source of natural antioxidants.

Type
Research Article
Copyright
Copyright © Proprietors of Journal of Dairy Research 2013 

Access options

Get access to the full version of this content by using one of the access options below. (Log in options will check for institutional or personal access. Content may require purchase if you do not have access.)

References

Cheison, SC, Zhang, W & Xu, SY 2007 Use of macroporous adsorption resin for simultaneous desalting and debittering of whey protein hydrolysates. International Journal of Food Science and Technology 42 12281239CrossRefGoogle Scholar
Cumby, N, Zhong, Y, Naczk, M & Shahidi, F 2008 Antioxidant activity and water-holding capacity of canola protein hydrolysates. Food Chemistry 109 144148CrossRefGoogle ScholarPubMed
Davalos, A, Miguel, M, Bartolome, B & Lopez-Fandino, R 2004 Antioxidant activity of peptides derived from egg white proteins by enzymatic hydrolysis. Journal of Food Protection 67 19391944CrossRefGoogle ScholarPubMed
Demers-Mathieu, V, Gauthier, SF, Britten, M, Fliss, I, Robitaille, G & Jean, J 2013 Antibacterial activity of peptides extracted from tryptic hydrolyzate of whey protein by nanofiltration. International Dairy Journal 28 94101Google Scholar
Elias, RJ, Kellerby, SS & Decker, EA 2008 Antioxidant activity of proteins and peptides. Critical Reviews in Food Science 48 430441Google Scholar
Guo, YX, Pan, DD & Tanokura, M 2009 Optimisation of hydrolysis conditions for the production of the angiotensin-1 converting enzyme (ace) inhibitory peptides from whey protein using response surface methodology. Food Chemistry 114 328333Google Scholar
Hogan, S, Zhang, L, Li, J, Wang, H & Zhou, K 2009 Development of antioxidant rich peptides from milk protein by microbial proteases and analysis of their effects on lipid peroxidation in cooked beef. Food Chemistry 117 438443Google Scholar
Kim, SB & Lim, JW 2004 Calcium-binding peptides derived from tryptic hydrolysates of cheese whey protein. Asian Australian Journal of Animal Sciences 17 14591464CrossRefGoogle Scholar
Korhonen, H & Pihlanto, A 2006 Bioactive peptides: production and functionality. International Dairy Journal 16 945960Google Scholar
Li, B, Chen, F, Wang, X, Ji, B & Wu, Y 2007 Isolation and identification of antioxidative peptides from porcine collagen hydrolysate by consecutive chromatography and electrospray ionization-mass spectrometry. Food Chemistry 102 11351143CrossRefGoogle Scholar
Lu, RR, Qian, P, Sun, Z, Zhou, XH, Chen, TP, He, JF, Zhang, H & Wu, JP 2010 Hempseed protein derived antioxidative peptides: purification, identification and protection from hydrogen peroxide-induced apoptosis in pc12 cells. Food Chemistry 123 12101218CrossRefGoogle Scholar
Mitsuta, H, Yasumoto, K & Iwami, K 1996 Antioxidative action of indole compounds during the autoxidation of linoleic acid. Eiyo to Shokuryo 19 210214CrossRefGoogle Scholar
Moure, A, Dominguez, H & Parajo, JC 2006 Antioxidant properties of ultrafiltration-recovered soy protein fractions from industrial effluents and their hydrolysates. Process Biochemistry 41 447456Google Scholar
Osawa, T & Namiki, M 1985 Natural antioxidant isolated from eucalyptus leaf waxes. Journal of Agricultural and Food Chemistry 33 770780Google Scholar
Park, P-J, Jung, W-K, Nam, K-S, Shahidi, F & Kim, S-K 2001 Purification and characterization of antioxidative peptides from protein hydrolysate of lecithin-free egg yolk. Journal of American Oil Chemist's Society 78 651656Google Scholar
Parrado, J, Miramontes, E, Jover, M, Gutierrez, JF, Collantes de Teran, L & Bautista, J 2006 Preparation of a rice bran enzymatic extract with potential use as functional food. Food Chemistry 98 742748CrossRefGoogle Scholar
Pea-Ramos, EA & Xiong, YL 2001 Antioxidative activity of whey protein hydrolysates in a liposomal system. Journal of Dairy Science 84 25772583Google Scholar
Pena-Ramos, EA & Xiong, YL 2002 Antioxidant activity of soy protein hydrolysates in a liposomal system. Journal of Food Science 67 29522956CrossRefGoogle Scholar
Peng, XY, Xiong, YLL & Kong, BH 2009 Antioxidant activity of peptide fractions from whey protein hydrolysates as measured by electron spin resonance. Food Chemistry 113 196201Google Scholar
Rajapakse, N, Mendis, E, Jung, W-K, Je, J-Y & Kim, S-K 2005 Purification of a radical scavenging peptide from fermented mussel sauce and its antioxidant properties. Food Research International 38 175182Google Scholar
Ramos, OL, Santos, AC, Leao, MV, Pereira, JO, Silva, SI, Fernandes, JC, Franco, MI, Pintado, ME & Malcata, FX 2012 Antimicrobial activity of edible coatings prepared from whey protein isolate and formulated with various antimicrobial agents. International Dairy Journal 25 132141Google Scholar
Ren, J, Zhao, M, Shi, J, Wang, J, Jiang, Y, Cui, C, Kakuda, Y & Xue, SJ 2008 Purification and identification of antioxidant peptides from grass carp muscle hydrolysates by consecutive chromatography and electrospray ionization-mass spectrometry. Food Chemistry 108 727736Google Scholar
Rui, XU 2009 Calcium binding of peptides derived from enzymatic hydrolysates of whey protein concentrate. International Journal of Dairy Technology 62 170173CrossRefGoogle Scholar
Saiga, A, Tanabe, S & Nishimura, T 2003 Antioxidant activity of peptides obtained from porcine myofibrillar proteins by protease treatment. Journal of Agricultural and Food Chemistry 51 36613667Google Scholar
Shimizu, M 2004 Food-derived peptides and intestinal functions. Biofactors 21 4347CrossRefGoogle ScholarPubMed
Unal, G & Akalin, AS 2012 Antioxidant and angiotensin-converting enzyme inhibitory activity of yoghurt fortified with sodium calcium caseinate or whey protein concentrate. Dairy Science and Technology 92 627639CrossRefGoogle Scholar
Xu, R, Liu, N, Xu, X & Kong, B 2011 Antioxidative effects of whey protein on peroxide-induced cytotoxicity. Journal of Dairy Science 94 37393746CrossRefGoogle ScholarPubMed
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. Biotechnology Letters 34 20012006Google Scholar