Protective Effects of Novel Antioxidant Peptide Purified from Alcalase Hydrolysate of Velvet Antler Against Oxidative Stress in Chang Liver Cells in Vitro and in a Zebrafish Model In Vivo.

Ding Ding,Lee Lee,Ko Ko,Moon Moon

doi:10.3390/ijms20205187

Ding Ding, Lee Lee + Show 2 more

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https://doi.org/10.3390/ijms20205187

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Abstract

Velvet antler has a long history in traditional medicine. It is also an important healthy ingredient in food as it is rich in protein. However, there has been no report about antioxidant peptides extracted from velvet antler by enzymatic hydrolysis. Thus, the objective of this study was to hydrolyze velvet antler using different commercial proteases (Acalase, Neutrase, trypsin, pepsin, and α-chymotrypsin). Antioxidant activities of different hydrolysates were investigated using peroxyl radical scavenging assay by electron spin resonance spectrometry. Among all enzymatic hydrolysates, Alcalase hydrolysate exhibited the highest peroxyl radical scavenging activity. Alcalase hydrolysate was then purified using ultrafiltration, gel filtration, and reverse-phase high performance liquid chromatography. The purified peptide was identified to be Trp-Asp-Val-Lys (tetrapeptide) with molecular weight of 547.29 Da by Q-TOF ESI mass spectroscopy. This purified peptide exhibited strong scavenging activity against peroxyl radical (IC50 value, 0.028 mg/mL). In addition, this tetrapeptide showed significant protection ability against AAPH-induced oxidative stress by inhibiting of reactive oxygen species (ROS) generation in Chang liver cells in vitro and in a zebrafish model in vivo. This research suggests that the tetrapeptide derived from Alcalase-proteolytic hydrolysate of velvet antler are excellent antioxidants and could be effectively applied as functional food ingredients and pharmaceuticals.

Highlights

Reactive oxygen species (ROS) are chemically reactive species containing oxygen
To the best of our knowledge, there have been no reports about antioxidant peptides extracted from velvet antler by enzymatic hydrolysis
Yields of velvet antler enzymatic hydrolysates measured by dry weight were observed to be 34.09%, 12.39%, 38.96%, 23.81%, and 29.75% for trypsin, pepsin, α-chymotrypsin, Neutrase, and Alcalase, respectively (Table 1)

Summary

Introduction

Reactive oxygen species (ROS) are chemically reactive species containing oxygen. ROS are normally produced in living organisms during metabolism of oxygen. Under normal conditions in our body, ROS can be effectively eliminated by antioxidant defense systems such as endogenous antioxidant enzymes and non-enzymatic factors [1]. Overproduction of ROS by various factors can cause oxidative stress and lead to a variety of pathological conditions, including metabolic impairments such as inflammation, aging, cancer, and cardiovascular diseases [2]. Sufficient amount of antioxidants need to be consumed to prevent or slow down oxidative stress induced by ROS. The amount of synthetic antioxidants used by humans is under strict regulation due to their potential health hazards [3]. Natural antioxidants without side effects or toxicity have attracted great interest

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