Isolation,identification and activity evaluation of Mytilus coruscus antioxidant peptides
Objective To prepare antioxidant peptides from Mytilus coruscus,using enzymatic hydrolysis and modern separation and purification technology.Methods The scavenging rates of DPPH radical(DPPH·),hydroxyl radical(HO·),and 2,2'-biazo-bis-3-ethylbenzothiazoline-6-sulfonic acid free radical(ABTS·)were measured as the index to optimize the conditions of enzymatic hydrolysis.Antioxidant peptides were isolated by ultrafiltration,gel filtration chromatography,and reverse high-performance liquid chromatography,and their antioxidant activities were evaluated.Results The optimal conditions for enzymatic hydrolysis were as follows:papain,enzymatic hydrolysis time of 3.0 h,enzyme dosage of 3.0％,feed-liquid ratio of 1∶50.Four peptides were isolated and identified as HK1:Gln-Thr-Asp(362.34 Da);HK2:His-Gln-Glu-Glu(541.52 Da);HK3:Leu-Glu-Gly-Asp-Thr(533.54 Da);HK4:Thr-Gln-Glu(376.37 Da)by reverse high-performance liquid chromatography and amino acid N-terminal sequencing.Their EC50 values for DPPH·scavenging were(0.523±0.066),(0.419±0.042),(0.467±0.078),and(0.682±0.134)mg/mL,respectively;and their EC50 values for HO·scavenging were(0.587±0.150),(0.866±0.123),(1.009±0.060),(1.014±0.122)mg/mL,respectively;their EC50 values for O2-·scavenging were(0.239±0.021),(0.245±0.019),(0.611±0.012),(0.716±0.024)mg/mL,respectively;the EC50 values of ABTS·scavenging were(0.218±0.109),(0.117±0.013),(0.121±0.019),and(0.415±0.054)mg/mL,respectively.Conclusion This study concluded that enzymatic hydrolysis of Mytilus coruscus protein by papain could produce peptides with significant free radical scavenging activities which could provide a theoretical basis and reference for the development and utilization of Mytilus coruscus.
Mytilus coruscushydrolyzed peptidesantioxidant activityisolation and purification
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