title: Anti-Inflammatory and Antioxidant Properties of Casein Hydrolysate Produced Using High Hydrostatic Pressure Combined with Proteolytic Enzymes.
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.
authors.affiliation: Centre for Pharmacy & Health Research, Faculty of Pharmacy and Pharmaceutical Sciences, University of Alberta, 11361-87 Ave, Edmonton, AB T6G 2E1, Canada. bamdad@ualberta.ca.
authors.forename: Fatemeh
authors.initials: F
authors.lastname: Bamdad
authors.affiliation: Centre for Pharmacy & Health Research, Faculty of Pharmacy and Pharmaceutical Sciences, University of Alberta, 11361-87 Ave, Edmonton, AB T6G 2E1, Canada. sshin1@ualberta.ca.
authors.forename: Seulki Hazel
authors.initials: SH
authors.lastname: Shin
authors.affiliation: Center for Nutraceutical and Pharmaceutical Materials, Myongji University, Yongin, Gyeonggi 449-728, Korea. jwsuh@mju.ac.kr.
authors.forename: Joo-Won
authors.initials: JW
authors.lastname: Suh
authors.affiliation: Centre for Pharmacy & Health Research, Faculty of Pharmacy and Pharmaceutical Sciences, University of Alberta, 11361-87 Ave, Edmonton, AB T6G 2E1, Canada. nimalara@ualberta.ca.
authors.forename: Chamila
authors.initials: C
authors.lastname: Nimalaratne
authors.affiliation: Centre for Pharmacy & Health Research, Faculty of Pharmacy and Pharmaceutical Sciences, University of Alberta, 11361-87 Ave, Edmonton, AB T6G 2E1, Canada. jwsuh@mju.ac.kr.
authors.forename: Hoon
authors.initials: H
authors.lastname: Sunwoo
mesh_terms: D000818:Animals; D000893:Anti-Inflammatory Agents / Q000138:chemical synthesis* / Q000494:pharmacology*; D000975:Antioxidants / Q000138:chemical synthesis* / Q000494:pharmacology*; D002364:Caseins / Q000138:chemical synthesis* / Q000494:pharmacology*; D002460:Cell Line; D002470:Cell Survival / Q000187:drug effects; D016207:Cytokines / Q000096:biosynthesis; D016166:Free Radical Scavengers / Q000138:chemical synthesis / Q000494:pharmacology; D006863:Hydrogen-Ion Concentration; D006868:Hydrolysis; D006874:Hydrostatic Pressure; D018836:Inflammation Mediators / Q000378:metabolism; D007502:Iron Chelating Agents / Q000138:chemical synthesis / Q000494:pharmacology; D008264:Macrophages / Q000187:drug effects / Q000276:immunology / Q000378:metabolism; D051379:Mice; D008970:Molecular Weight; D010447:Peptide Hydrolases; D013997:Time Factors
chemical_list: D000893:Anti-Inflammatory Agents; D000975:Antioxidants; D002364:Caseins; D016207:Cytokines; D016166:Free Radical Scavengers; D018836:Inflammation Mediators; D007502:Iron Chelating Agents; C475729:casein hydrolysate; D010447:Peptide Hydrolases
keywords: anti-inflammation; antioxidant activity; casein; enzymatic hydrolysis; high hydrostatic pressure