Anti-Inflammatory and Antioxidant Properties of Peptides Released from β-Lactoglobulin by High Hydrostatic Pressure-Assisted Enzymatic Hydrolysis

Abstract

Background: β-lactoglobulin hydrolysates (BLGH) have shown antioxidant, antihypertensive, antimicrobial, and opioid activity. In the current study, an innovative combination of high hydrostatic pressure and enzymatic hydrolysis (HHP–EH) was used to increase the yield of short bioactive peptides, and evaluate the anti-inflammatory and antioxidant properties of the BLGH produced by the HHP–EH process. Method: BLG was enzymatically hydrolyzed by different proteases at an enzyme-to-substrate ratio of 1:100 under HHP (100 MPa) and compared with hydrolysates obtained under atmospheric pressure (AP-EH at 0.1 MPa). The degree of hydrolysis (DH), molecular weight distribution, and the antioxidant and anti-inflammatory properties of hydrolysates in chemical and cellular models were evaluated. Results: BLGH obtained under HHP–EH showed higher DH than the hydrolysates obtained under AP-EH. Free radical scavenging and the reducing capacity were also significantly stronger in HHP-BLGH compared to AP-BLGH. The BLGH produced by alcalase (Alc) (BLG-Alc) showed significantly higher antioxidant properties among the six enzymes examined in this study. The anti-inflammatory properties of BLG-HHP-Alc were observed in lipopolysaccharide-stimulated macrophage cells by a lower level of nitric oxide production and the suppression of the synthesis of pro-inflammatory cytokines. Peptide sequencing revealed that 38% of the amino acids in BLG-HHP-Alc are hydrophobic and aromatic residues, which contribute to its anti-inflammatory properties. Conclusions: Enzymatic hydrolysis of BLG under HHP produces a higher yield of short bioactive peptides with potential antioxidant and anti-inflammatory effects.

Comparison of antioxidant activities of bovine whey proteins before and after simulated gastrointestinal digestion

“…Alcalase and neutrase hydrolysates from whey proteins also protected lung fibroblast and umbilical vein endothelial cells by boosting intracellular antioxidant defenses (Kong et al, 2012;O'Keeffe and FitzGerald, 2014). In fact, the hydrolysis of β-LG with alcalase produced the most antioxidant fraction containing the peptides f(27-38) DIQKVAGTWYSL, f(33-38) GTWYSL, f(39-48) AMAASDISLL, f(40-48) MAASDISLL, f(61-73) ELKPTPEGDLEIL, f(87-98) IIAEKTKIPAVF, f(112-121) DTDYKKYLLF, and f(165-172) LSFNPTQL (Bamdad et al, 2017). It is important to note that antioxidant activities from commercial enzyme hydrolysates in our study are likely to be affected by the standard thermal step required for enzyme inactivation, which is known to affect protein denaturation and aggregation (Joyce et al, 2018).…”

“…In addition, whey fractions with a molecular weight below 5 kDa consistently exert better antioxidant properties than higher-molecular weight fractions (de Castro and Sato, 2014;Liu et al, 2014;O'Keeffe and FitzGerald, 2014). Recently, Bamdad et al (2017) identified 8 peptides present in the most antioxidant fraction of β-LG hydrolyzed by alcalase.…”

Production and processing of antioxidant bioactive peptides: A driving force for the functional food market

“…Bamdad et al (2017b) reported that enzyme hydrolysis of casein protein under HHP at 100 MPa improved the degree of hydrolysis and antioxidant properties compared to 200 MPa and atmospheric pressure enzyme hydrolysis. Similarly, Bamdad et al (2017a) revealed that HHP assisted enzyme hydrolysis of β-lactoglobulin protein at 100 MPa resulted in higher degree of hydrolysis, and enhanced free radical scavenging and reducing capacity than the hydrolysates produced with the same enzymes at atmospheric pressure. The most recent study by Dong et al (2019) also demonstrated that the degree of hydrolysis, and reducing power and DPPH radical scavenging capacity of peanut protein hydrolysate treated at 300 MPa for 60 min and 100 MPa for 180 min are significantly higher than untreated and 500 MPa treated proteins.…”

Advances on Food-Derived Peptidic Antioxidants—A Review

“…HP pretreatment/auxiliary treatment can increase the hydrolysis rate and DH of the substrate protein, promote the release of low Mw peptides and essential amino acids, and improve the antioxidant activity of the hydrolysate [106], which has been applied to promote the proteolysis process of legume [107,108] and peanut protein [109]. Additionally, high hydrostatic pressure (HHP) also has the ability to improve the efficiency of enzymatic hydrolysis [110]. HHP auxiliary treatment can improve the enzymatic hydrolysis efficiency of Corolase PP, reduce the surface hydrophobicity of the hydrolysate, and increase the production of small peptides [111,112].…”