Novel Antioxidant Peptides Purified from Mulberry (Morus atropurpurea Roxb.) Leaf Protein Hydrolysates with Hemolysis Inhibition Ability and Cellular Antioxidant Activity

Abstract

Neutrase-hydrolysates hydrolyzed from mulberry leaf proteins were separated by ion exchange chromatography, gel filtration chromatography, and semipreparative reverse-phase HPLC. Purified fractions were analyzed for their radical scavenging activity, hemolysis inhibition ability, and cellular antioxidant activity (CAA). Three new antioxidant peptides, P1 (SVL, 317 Da), P2 (EAVQ, 445 Da), and P3 (RDY, 452 Da), were obtained from the most active HPLC fraction (R1) and identified using UPLC-QTOF-MS. These three peptides were then synthesized, and their antioxidant activities were analyzed. P1 and P2 had no ability to inhibit hemolysis of erythrocytes but did show antioxidant activity on HepG2 cells. P3 showed the highest hemolysis inhibition ability (92%) and CAA value (2204 μM QE/100 g peptide). The Tyr residues at the C-terminal region play an important role in the antioxidant activity in P3. Thus, the natural peptide R1 and synthesized P3 could be used as antioxidants and might be promising components of functional foods.

Advances on Food-Derived Peptidic Antioxidants—A Review

“…Cuttlefish-purified peptides can obviously inhibit the oxidation process of linoleic acid, effectively protect the DNA damage induced by • OH radicals, and have no toxic effect to cells at higher concentrations [ 159 ]. In addition, some studies have confirmed that PAs have certain potential in the treatment of cancer [ 275 ], malaria [ 276 ], hypertension [ 277 , 278 ] and blood diseases [ 224 ], and are expected to become therapeutic drugs. Bioactive peptides derived from rapeseed protein (LY, RALP, GHS) can significantly inhibit the secretion of NO, il-6 and tumor necrosis factor stimulated by lipopolysaccharide, and then repair the damage of spontaneously hypertensive cells caused by oxidative stress, which has potential application value in protecting the body from oxidative and inflammatory damage [ 279 ].…”

Preparation and Evaluation of Gallate Ester Derivatives Used as Promising Antioxidant and Antibacterial Inhibitors

“…The RSA of the gallate ester derivatives against the stable DPPH ⋅ was determined spectrophoto‐ metrically [39,40] . The methods of Casimir and Wang et al .…”

Cellular antioxidant effect of bioactive peptides and molecular mechanisms underlying: beyond chemical properties

“…In the majority of cases, the common cell lines used include: human epithelial colorectal adenocarcinoma (Caco-2) (Zhang, Tong, et al, 2019;Tonolo et al, 2020), human liver carcinoma (HepG2) (Chen et al, 2019;Sun et al, 2019), human normal hepatoma (Chang) (Je et al, 2015;Ding et al, 2019), cervical adenocarcinoma (HeLa) (Ma et al, 2016;Mart ınez-Leo et al, 2019), human umbilical vein endothelial (HUVEC) (Liang et al, 2018;Xu et al, 2018), human neuroblastoma (SH-SY5Y) (Lee & Hur, 2019;Sheng et al, 2019), rat adrenal pheochromocytoma (PC12) (Lee & Hur, 2019;Liu et al, 2019) and mouse macrophage (RAW 264.7) (Saisavoey et al, 2016;Indiano-Romacho et al, 2019) cells. Also, the most common inductor of oxidative stress used in the above-mentioned studies was hydrogen peroxide (H 2 O 2 ), a reactive oxygen species (ROS), because it can rapidly diffuse across cell membranes and can trigger programmed cell death (Di Marzo et al, 2018).…”

“…Yang, Li, et al (2017) purified two peptides (KWFCT and Ac-QWFCT) from 3-10 kDa peptide fraction from pine nut (Pinus koraiensis) meal protein hydrolysate with cellular antioxidant activity (612.8 and 916.3 µM quercetin equivalents/100 g, respectively). Similarly, Sun et al (2019) synthesised three peptides (SVL, EAVQ and RDY) derived from Mulberry (Morus atropurpurea Roxb.) leaf protein hydrolysate that exhibited cellular antioxidant activity (1706, 1501 and 2204 lM quercetin equivalents/100 g peptide, respectively).…”