Abstract
Casein-derived peptides are shown to possess radical scavenging and metal chelating properties. The objective of this study was to evaluate novel anti-inflammatory properties of casein hydrolysates (CH) produced by an eco-friendly process that combines high hydrostatic pressure with enzymatic hydrolysis (HHP-EH). Casein was hydrolysed by different proteases, including flavourzyme (Fla), savinase (Sav), thermolysin (Ther), trypsin (Try), and elastase (Ela) at 0.1, 50, 100, and 200 MPa pressure levels under various enzyme-to-substrate ratios and incubation times. Casein hydrolysates were evaluated for the degree of hydrolysis (DH), molecular weight distribution patterns, and anti-inflammatory properties in chemical and cellular models. Hydrolysates produced using HHP-EH exhibited higher DH values and proportions of smaller peptides compared to atmospheric pressure-enzymatic hydrolysis (AP-EH). Among five enzymes, Fla-digested HHP-EH-CH (HHP-Fla-CH) showed significantly higher antioxidant properties than AP-Fla-CH. The anti-inflammatory properties of HHP-Fla-CH were also observed by significantly reduced nitric oxide and by the suppression of the synthesis of pro-inflammatory cytokines in lipopolysaccharide (LPS)-stimulated RAW 264.7 macrophage cells. Liquid chromatography with tandem mass spectrometry (LC-MS/MS) revealed that 59% of the amino acids of the peptides in HHP-Fla-CH were composed of proline, valine, and leucine, indicating the potential anti-inflammatory properties. In conclusion, the HHP-EH method provides a promising technology to produce bioactive peptides from casein in an eco-friendly process.
Highlights
It is well-recognized that food-derived peptides can exert beneficial biological activities in addition to their basic nutritional role [1]
1:50) improved the degree of hydrolysis (DH) and resulted in hydrolysates with improved antioxidant properties compared to atmospheric pressure hydrolysis
The molecular weight distribution of hydrolysates reflected in SEC–HPLC and MALDI–TOF analysis confirmed that the high hydrostatic pressure with enzymatic hydrolysis (HHP-EH) method produces a greater proportion of short peptides compared to atmospheric pressure-enzymatic hydrolysis (AP-EH)
Summary
It is well-recognized that food-derived peptides can exert beneficial biological activities in addition to their basic nutritional role [1]. 2–20 amino acids) and may possess antioxidant, anti-inflammatory, antihypertensive, antimicrobial, and anticancer properties that have a potential role in maintaining or promoting human health [2]. Both anti-inflammatory and antioxidant properties are two of the main topics considered for preventing chronic diseases such as cardiovascular disease and cancer due to oxidative stress, as well as abnormal inflammatory responses. Casein phosphopeptides derived from gastrointestinal and commercial proteases may act as multifunctional bioactive peptides with radical scavenging and metal chelating properties, and may play a role in enhancing mineral bioavailability [7]
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