Double enzyme hydrolysis for producing antioxidant peptide from egg white: Optimization, evaluation, and potential allergenicity

Abstract

Egg white is a good source of high-quality proteins in food products and an excellent source of antioxidant active peptides through hydrolysis. The hydrolysis conditions for the preparation of potent antioxidant peptides from egg white with chymotrypsin and pepsin were optimized by the response surface methodology. The antioxidant activity and potential allergenicity of the prepared peptides were evaluated by the oxidative damage model and IgE-binding capability, respectively. After ultrafiltration, the peptide produced using the optimized parameters (preheating time of 3.16 hr, hydrolysis time of 3 hr, a sample/solution ratio of 10%, multiple enzymes ratio (E1/E2) of 1.7:1, and E/S of 0.4%) showed antioxidant activity of was 30.86 μmol AAE/g DW and with low potential for allergenicity. The optimized method is efficient and economical and may be applied to the industrial production of antioxidant peptides to obtain nutraceutical and pharmaceutical agents with low sensitivity. However, further in vivo studies must be conducted. Practical applicationsEgg is consumed as an excellent source of protein. Antioxidant peptides released from egg white is considered to be used in food preservation and human health.Few researches on the optimization of egg peptides were aimed to obtain practical techniques used in the food industry. In this paper, egg white hydrolysis peptide with high antioxidant property and low potential allergenicity was prepared after the optimization of double enzyme hydrolysis. The products could be a natural health care products derived from a dietary source and considered using in additive during food production and health food. And the methods used are economical and energysaving and could be developed to utilize in the food industry. K E Y W O R D S antioxidant peptide, egg white, hydrolysis, optimization S U PP O RTI N G I N FO R M ATI O N Additional supporting information may be found online in the Supporting Information section. How to cite this article: Yuan J, Zheng Y, Wu Y, Chen H, Tong P, Gao J. Double enzyme hydrolysis for producing antioxidant peptide from egg white: Optimization, evaluation, and potential allergenicity. J Food Biochem. 2020;44:e13113.

Advances on Food-Derived Peptidic Antioxidants—A Review

“…At present, due to the lack of animal model experiments and human clinical trials, the research progress of antioxidant peptides in terms of bioavailability is minimal, which limits their commercialization process. On the other hand, in the application process of the food matrix, the influence of food ingredients and processing conditions on the bioavailability of PAs, as well as the production cost, biological activity, efficacy and safety of peptide-containing foods should be considered [280][281][282]. Before the development of large-scale foods containing PAs, it must be confirmed that the addition of these peptides will not cause negative effects and still retain their antioxidant activity.…”

Can food processing produce hypoallergenic egg?

“…By simulating gastrointestinal digestion in vitro, Martos et al (2013) found that egg white hydrolysis was promoted in the presence of egg yolk, but the presence of yolk components did not further reduce its IgE-binding capacity after hydrolysis. Apart from the hydrolysis induced by simulated gastrointestinal digestion, the IgE binding of egg white proteins has also been reported to be reduced by using individual enzymes, such as alcalase, chymotrypsin, and pepsin (Yang, Long et al, 2017;Yuan et al, 2019). Emulsification increased the digestibility of egg and further reduced its IgE-binding capacity (Jiménez-Saiz, Ruiz-Henestrosa, et al, 2012).…”

“…Reduced IgE-binding capacity Yuan et al (2019) Egg white proteins were preheated at 85°C for 30 min, then Alcalase (free or immobilized) was used for hydrolysis at pH 8 at 60°C from 10 to 250 min Reduced IgE-binding capacity Yang, Long et al (2017) Egg white in the presence or absence of egg yolk 2014; Hildebrandt et al (2008) antioxidant activity (Yang, Tu, Wang, Zhang, & Song, 2018). Both interactions with epigallo-catechin 3-gallate and chlorogenic acid reduced IgE-binding capacity of ovalbumin, due to the shielding of the linear IgE epitope (Lu et al, 2018;Ognjenović et al, 2014).…”

Chicken Egg Proteins and Derived Peptides with Antioxidant Properties

“…Commercial enzymes are preferred over naturally occurring ones because their specific characteristics such as optimal pH, temperature or cleavage site are well defined [40]. Normally, one or several proteases are used to obtain proteins hydrolysate [41] being pepsin, trypsin, alcalase and papain some of the most popular enzymes used [12,[42][43][44]. Alternatively, non-commercial enzymes have also applied to produce functional egg peptides to reduce the cost of hydrolysis [41,45].…”

“…A wide screening of the conditions required to obtain the pepsin hydrolysates with the strongest antioxidant capacity was done by Lin et al which determined that 4.56% of egg white as starting material with an enzyme to substrate ratio of 1.58% at pH 1.99 and 37 • C for 1 h were the optimal conditions, although they do not specify the activity of the pepsin used and a pre-treatment of 10 min at 90 • C was applied to denature egg white proteins [12]. Other studies have also probed the antioxidant capacity of hydrolyzed egg white with pepsin using other methods such as Ferric Reducing Antioxidant Power (FRAP) assay [43] or measurement of oxidative stress inhibitory activity in cell lines such as in the study Garcés-Rimón et al, where they observed a dose-dependent inhibition of reactive oxygen species production in a macrophage cell line after the treatment with an egg white hydrolysate obtained with pepsin [64].…”