Pinna pectinata was used as raw material to extract and the antioxidant levels of the hydrolyzed peptides and the protective effects against H2O2-induced oxidative damage were explored.The controlled enzymatic hydrolysis technology was used to screen the best hydrolase and optimize the enzymatic hydrolysis conditions, DPPH radical scavenging rates and the degrees of hydrolysis were used as the main detection indexes.Ultrafiltration, Q sepharose fast flow (QFF) anion column, reversed phase high-performance liquid chromatography (RP-HPLC), amino acid N-terminal sequencing, and mass spectrometry were used to isolate and identify active enzymatic hydrolyzed peptides.H2O2-induced human umbilical vein endothelial cells (HUVECs) were used as an oxidative damage model to study the protective effects of active peptides against oxidative damage.The optimal enzymolysis conditions were: papain, 37 ℃, pH 8.0, enzyme dosage of 3.0%, solid-liquid ratio of 1∶20, enzymolysis time of 3.0 h.Five oligopeptides were isolated and identified from the enzymolysis products, which were identified by mass spectrometry and amino acid sequence analysis as Val-Ala-His (JY1, 325.18 Da), Glu-Pro (JY2, 244.11 Da), Ala-Val (JY3, 188.12 Da), Trp-Tyr (JY4, 367.15 Da), and Trp-Val (JY5, 303.16 Da).Of which, JY1 and JY4 showed better antioxidant activities, and the EC50 values against DPPH radicals, ABTS cationic radicals, ·O2- were (2.102±0.01), (4.215±0.024), (3.019±0.021) mg/mL, (2.981±0.014), (5.385±0.027), (4.204±0.031) mg/mL, respectively.They both had significant protective effects on H2O2-induced HUVECs damage, and can significantly improve cell survival rates and reduce cell damage.The enzymatic hydrolysis of P.pectinata protein have antioxidant activities and can protect HUVECs from H2O2 induced oxidative damage.The results of this study can provide theoretical basis and reference for the prevention and treatment of cardiovascular diseases, the development of antioxidant products, and utilization of P.pectinata.
LI Zhen
,
JIN Danqing
,
XU Xuan
,
WANG Kun
,
LI Jiayao
,
REN Chunzhi
,
TANG Hongjun
,
WANG Yumei
,
SUN Kunlai
. Preparation of antioxidant peptides from Pinna (Atrina) pectinata and their protective effects on oxidative damage of human umbilical vein endothelial cells[J]. Food and Fermentation Industries, 2024
, 50(23)
: 209
-217
.
DOI: 10.13995/j.cnki.11-1802/ts.038450
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