Purification and identification of dual-enzyme hydrolysates obtained from defatted walnut and its antioxidant effects on
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ORIGINAL PAPER
Purification and identification of dual‑enzyme hydrolysates obtained from defatted walnut and its antioxidant effects on d‑galactose‑induced aging mice Qi Jia1 · Jin‑Fang Yuan1 · Hui‑Ping Liu1 · Meng‑Yu Li1 · Ya‑Ru Wu1 Received: 6 May 2020 / Accepted: 12 October 2020 © Springer Science+Business Media, LLC, part of Springer Nature 2020
Abstract The purpose of this study was to explore the antioxidant activity in vivo of dual-enzyme hydrolysates obtained from defatted walnut, and to purify and identify it. The walnut protein hydrolysates (WPH) were obtained through two-step enzymatic hydrolysis of alkaline protease and trypsin. The effects of WPH on the oxidative stress-related biochemical indexes of d-galactose-treated mice were investigated. Next, WPH was passed through a Sephadex G-25 gel chromatography, anion exchange chromatography and Sephadex G-15 gel chromatography. Subsequently, peptides in the high antioxidant activity fraction were identified by RP-nano-HPLC–MS/MS. Finally, 14 potential peptides of length 11 AAs to 21 AAs with antioxidant activity were identified from WPH. Keywords Walnut protein · Antioxidant peptides · Aging mice · Chromatographic separation · Amino acid sequence
Introduction Protein hydrolysates added to food may protect the food from lipid oxidation (and oxidation of other macromolecules) by physical or chemical means [1]. The production of peptides through hydrolytic reactions seems to be the most promising technique to form proteinaceous antioxidants since peptides have substantially higher antioxidant activity than intact proteins [2], such as egg white protein hydrolysate [3], rice bran protein hydrolysates [4], Sea cucumber hydrolysates [5], tilapia skin collagen polypeptide [6], amaranth proteins hydrolysate [7], feather keratin hydrolysates [8, 9], whey protein hydrolysates [9, 10], fish protein hydrolysates [11–15]. These peptides usually contain 3–20 amino acid residues, and their activity depends on their amino acid composition, structure and hydrophobicity [3, 16–19]. * Hui‑Ping Liu [email protected] 1
State Key Laboratory of Food Nutrition and Safety, Key Laboratory of Food Nutrition and Safety, Ministry of Education of China, College of Food Science and Engineering, Tianjin University of Science and Technology, No. 29, 13th Road, Tianjin Economic and Technical Development Zone, Tianjin 300457, People’s Republic of China
Walnut (Juglans regain L) is one of the four major nuts. The walnut kernel has various biological activities, such as antiproliferative activity [20], antibacterial [21], anti-aging, immune regulation and so on. Shanxi paper-shell walnut Jinlong No.1 has the following characteristics: the nut is nearly round, the fruit base is slightly concave, the fruit top is flat; The shell surface is smooth, with small pitting points and light color. The suture line is narrow and flat, and the shell thickness is 1.1 mm. The color of walnut meal is light and fragrant. The fat content is 64.9% and the protein content is 14.32% [22]. At presen
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