Eur. J. Lipid Sci. Technol.

2008, 110, 489490

DOI 10.1002/ejlt.200800120

Editorial

489

Editorial Antioxidant activity of protein hydrolysates

The findings of numerous studies have confirmed that protein hydrolysates possess antioxidant activity. Hence, enzymatically-modified proteins may serve as natural sources of antioxidants in food models to preserve these foods and to extend their shelf-life. The properties of the hydrolysates dependent on a number of factors including the type of protein and enzyme employed the degree of hydrolysis (DH), the environmental conditions, and any substrate pretreatment. The capability of peptides to inhibit deleterious changes during lipid oxidation also depends on the presence of certain amino acid residues in the peptides. The main players here include tyrosine, histidine, methionine, and tryptophan. These amino acids are also capable of chelating pro-oxidant metal ions. The sources of protein utilized in the production of hydrolysates, which possess antioxidant activity, include soybean, canola, maize zein, egg white and yolk albumin, casein, whey, fish (e.g., tilapia, capelin), and pork [19]. Several methods to assess the antioxidant properties of hydrolysates so prepared have been employed and include a bulk oil system, emulsion system, liposomal system, meat/food model system with measurements of peroxide value, 2-thiobarbituric acid reactive substances (TBARS), antiradical activity against the DPPH and hydroxyl radicals, superoxide-scavenging activity, and ferric reducing/antioxidant power (FRAP) [19]. Results of experiments have indicated that the antioxidant properties of the hydrolysates dependent on the enzymes employed during hydrolysis. For example, hydrolysates prepared from canola protein using Flavourzyme (i.e., both with endo- and exo-peptidase activities) showed the highest antioxidant activity among all samples, whereas hydrolysates prepared with a combination of Alcalase (i.e., an endopeptidase) and Flavourzyme did not differ in antioxidant effectiveness from those generated by Alcalase alone [6]. Hydrolysates prepared from whey protein isolate by pure enzyme (i.e., pepsin, papain, trypsin, chymotrypsin) treatments did not prevent TBARS formation in a liposomal system [3]. The antioxidant activity was noted in the case of hydrolysates prepared from WHI using commercial crude enzymes (i.e., proteases from Bacillus licheniformis, Aspergillus oryzae, as well as Bacillus protease complex). Under the influence of Alcalase (enzyme obtained from Bacillus licheniformis), an increase in activity for the reaction mixture was noted by the presence of cysteine-, methionine-, lysine-, and valine-containing protein fragments [5]. The antioxidant effect of these amino acid residues was reported long ago in a very important publication by Marcuse [10]. Based on a liposomal system involving maize zein hydrolysates, the lack of a direct relationship between antioxidant activity and %DH suggested that the presence of specific peptides liberated from protein and the ratio of key free amino acids are important factors when assessing the antioxidant activity of the hydrolysed protein. The results suggest a possible involvement of multiple reactions and processes leading to the inhibition of lipid oxidation; that is, the capability to donate a hydrogen atom, to stabilize or terminate radicals, and/or to chelate prooxidative metal ions. The peptide cleavages led to an enhanced Cu(II) binding by the opened carboxylic groups. The capability of different protein hydrolysates to chelate Fe(II) depends on the enzymes utilized in hydrolysis and the %DH of the hydrolysate [9]. In the case of emulsions and liposomes, peptides can also act as a physical barrier around fat droplets. According to Kong and Xiong [5], short peptides diffuse to the water-oil interface in a liposome system, where they can be adsorbed or loosely bind to the phospholipids membrane in the liposome where oxidation takes places. Protein hydrolysates can also protect meat against cholesterol oxidation. Enzymatically(EHC) and acid-derived hydrolysates of cracklings (AHC) prepared from collagenic sources demonstrated stability against cholesterol oxidation. EHC showed better inhibition against the formation of cholesterol oxidation products in stored meatballs. After 7 days of refrigerated storage, inhibition of in the formation of oxysterols in samples by added hydrolysates ranged from 29 to 54%. Butylated hydroxytoluene (BHT) was a weaker antioxidant than the hydrolysates against cholesterol oxidation in stored meatballs [11]. In a cholesteryl linoleate emulsion,

Ryszard Amarowicz

2008 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim

www.ejlst.com

490

Editorial

Eur. J. Lipid Sci. Technol. 2008, 110, 489490

which contained haematin and hydrogen peroxide as pro-oxidants, hydrolyzates prepared from cracklings limited the formation of hydroperoxides, conjugated dienes, and TBARS [12]. In lipid oxidation assays, enzymatically-derived Hydrolysate (CEH) exhibited superior antioxidative properties to those of acid-derived Hydrolysate (CAH); inhibition of peroxide formation by CEH ranged from 20 to 26%, whereas for CAH it ranged from 8 to 19% and depended on the time of incubation at 37 7C. The formation of conjugated dienes was inhibited by CEH by 8 to 40% and for CAH by 4 to18%. After 360 days of storage of pork meat meatballs, the content of oxysterols in samples treated with crackling hydrolysates was nearly 20% lower than that of the control sample [13]. The most toxic cholesterol, triol-cholesterol, was detected at the lowest concentration in all samples treated with the hydrolysates. When reflecting on the numerous publications reporting results pertaining to peptides and/ or hydrolysates as a source of natural antioxidants, it appears to be a hot interdisciplinary topic involving various aspects of lipid and protein science. In many laboratories, research on novel protein sources for hydrolysate preparations is in progress. Scientists are also trying to unravel or to provide an explanation for the complicated and difficult mechanisms dealing with the protection of lipid constituents against oxidation by peptides.

Ryszard Amarowicz Editor Division of Food Science, Institute of Animal Reproduction and Food Research, Polish Academy of Sciences, Olsztyn, Poland

References
[1] E. A. Pea-Ramos, Y. L. Xiong: Antioxidant activity of soy protein hydrolysates in a liposomal system. J Food Sci. 2002, 67, 29522956. [2] E.A. Pea-Ramos, Y. L. Xiong: Whey and soy protein hydrolysates inhibit lipid oxidation in cooked pork patties. Meat Sci. 2003, 64, 259263. [3] E.A. Pea-Ramos, Y. L. Xiong: Antioxidant activity of whey protein hydrolysates in a liposomal system. J Dairy Sci. 2001, 84, 25772583. [4] S. Sakanaka, Y. Tachibana: Active scavenging activity of egg-yolk protein hydrolysates and their effects on lipid oxidation in beef and tuna homogenates. Food Chem. 2006, 95, 243249. [5] B. Kong, Y. L. Xiong: Antioxidant activity of zein hydrolysates in a liposome system and the possible mode of action. J Agric Food Chem. 2006, 54, 60596068. [6] N. Cumby, Y. Zhong, M. Naczk, F. Shahidi: Antioxidant activity and water-holding capacity of canola protein hydrolysates. Food Chem. 2008, 109, 144148. [7] R. Amarowicz, F. Shahidi: Antioxidant activity of peptide fractions of capelin protein hydrolysates. Food Chem. 1997, 58, 355359. [8] F. Shahidi, R. Amarowicz (1996). Antioxidant activity of protein hydrolysates from aquatic species. J Am Oil Chem Soc. 1996, 73, 11971199. [9] S. Raghavan, H. G. Kristinsson: Antioxidant efficacy of alkali-treated tilapia protein hydrolysates: A comparison study of five enzymes. J Agric Food Chem. 2008, 56, 14341441. [10] R. Marcuse: Antioxidant effect of amino acids. Nature 1960, 186, 886887. [11] E. Flaczyk, M. Rudzinska, E. Wasowicz, J. Korczak, R. Amarowicz: Effect of cracklings hydro lysates on oxidative stability of pork meatballs fat. Food Res Int. 2006, 39, 924931. [12] E. Flaczyk, M. Rudzinska, J. Korczak, R. Amarowicz: Stabilization of cholesteryl linileate emulsion model system by cracklings hydrolysates. J Food Lipids. 2006, 13, 924931. [13] M. Rudzinska, E. Flaczyk, R. Amarowicz, E. Wasowicz, J. Korczak: Antioxidative effect of crackling hydrolysates during frozen storage of cooked pork meatballs. Eur Food Res Technol. 2007, 224, 293299. 2008 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim
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