Purification, structural analysis, and stability of antioxidant peptides from purple wheat bran
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RESEARCH ARTICLE
BMC Chemistry Open Access
Purification, structural analysis, and stability of antioxidant peptides from purple wheat bran Yan Zhao1,2*, Qi Zhao1,2 and Qingyu Lu1,2
Abstract Protein derived from purple wheat bran was hydrolyzed sequentially using alcalase proteases for the production of antioxidant peptides. Purple wheat bran protein (PWBP) hydrolysates were fractionated using size-exclusion (G-25) and ion-exchange chromatography methods to identify the structure of antioxidant peptides. The free radical scavenging activity of peptides purified from PWBP hydrolysates was evaluated using superoxide anion radical-scavenging activity and determination assays of Trolox equivalent antioxidant capacity (TEAC). Results demonstrated that purple wheat bran peptide F4-4 exhibited the highest antioxidant activity among other hydrolysates. F4-4 was further identified as Cys-Gly-Phe-Pro-Gly-His-Cys, Gln-Ala-Cys, Arg-Asn-Phe, Ser-Ser-Cys, and Trp-Phe by high performance liquid chromatography (HPLC) spectrometer coupled with Orbitrap Elite™ mass spectrometer (LC–MS/MS). Antioxidant peptides 2 and 4 showed improved stability when the temperature was lower than 80 °C. These peptides also demonstrated good digestive stability in vitro system by simulating gastrointestinal digestion. Keyword: Purple wheat bran, Antioxidant peptide, Purification, Structural analysis, Stability Introduction Antioxidants are substances that counteract the oxidation reaction caused by free radicals. Antioxidants are used to extend the shelf life of foods because they reduce lipid oxidation [1]. Synthetic antioxidants are commonly used in the food industry [2]. The most widely used synthetic antioxidants are butylated hydroxy anisole (BHA), butylated hydroxytoluene (BHT), and tert-butylhydroquinone (TBHQ). However, the use of synthetic antioxidants is strictly regulated because of their adverse effects to human health [3]. Studies have shown that these synthetic antioxidants can have varying degrees of toxicity in humans and affect the liver, spleen, and lung [4–6]. Therefore, the use of these synthetic chemical substances has been strictly limited [7]. Currently, considerable attention has been provided to natural antioxidants due
*Correspondence: [email protected] 1 National Engineering Laboratory of Wheat & Corn Further Processing, Henan University of Technology, Zhengzhou 450001, China Full list of author information is available at the end of the article
to their high efficacy and little or no side effects compared with synthetic antioxidants [8, 9]. Peptides derived from natural sources are natural antioxidants. These antioxidants have been the focus of research interest because of their potential health benefits associated with low molecular weight, low cost, high activity, functional properties, and safety [10, 11]. Many studies have produced bioactive peptides from plant protein sources. Thus, natural antioxidants from grains, vegetables, and plant substrates are crucial in protecting the body from reactive oxygen species (
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