Bioactive Peptides from Cartilage Protein Hydrolysate of Spotless Smoothhound and Their Antioxidant Activity In Vitro

Abstract

In the experiment, crude proteins from spotless smoothhound (Mustelus griseus), cartilages were isolated by HCl-Guanidine buffer, and its hydrolysate was prepared using trypsin at pH 8.0, 40 °C with a total enzyme dose of 2.5%. Subsequently, three antioxidant peptides were purified from the hydrolysate using membrane ultrafiltration, anion-exchange chromatography, gel filtration chromatography, and reverse phase high-performance liquid chromatography. The amino acid sequences of isolated peptides were identified as Gly-Ala-Glu-Arg-Pro (MCPE-A); Gly-Glu-Arg-Glu-Ala-Asn-Val-Met (MCPE-B); and Ala-Glu-Val-Gly (MCPE-C) with molecular weights of 528.57, 905.00, and 374.40 Da, respectively, using protein amino acid sequence analyzer and mass spectrum. MCPE-A, MCPE-B and MCPE-C exhibited good scavenging activities on 2,2-diphenyl-1-picrylhydrazyl radicals (DPPH•) (EC50 3.73, 1.87, and 2.30 mg/mL, respectively), hydroxyl radicals (HO•) (EC50 0.25, 0.34, and 0.06 mg/mL, respectively), 2,2′-azino-bis-3-ethylbenzothiazoline-6-sulfonic acid radicals (ABTS+•) (EC50 0.10, 0.05, and 0.07 mg/mL, respectively) and superoxide anion radicals (normalO2−•) (EC50 0.09, 0.33, and 0.18 mg/mL, respectively). MCPE-B showed similar inhibiting ability on lipid peroxidation with butylated hydroxytoluene (BHT) in a linoleic acid model system. Furthermore, MCPE-A, MCPE-B, and MCPE-C could protect H2O2-induced HepG2 cells from oxidative stress by decreasing the content of malonaldehyde (MDA) and increasing the levels of superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GSH-Px), and glutathione reductase (GSH-Rx). Glu, Gly, Met, and Pro in their sequences and low molecular weight could be attributed to the antioxidant activities of three isolated peptides. These results suggested that GAERP (MCPE-A), GEREANVM (MCPE-B), and AEVG (MCPE-C) from cartilage protein hydrolysate of spotless smoothhound might serve as potential antioxidants and be used in the pharmaceutical and health food industries.

High Fischer ratio oligopeptides determination from Antartic krill: Preparation, peptides profiles, and in vitro antioxidant activity

“…The EC 50 value of HFP was 4.90 mg/ml, which was lower than those of HFO from flaxseed (Udenigwe & Aluko, ), and anti‐oxidative oligopeptides from protein hydrolysates of weatherfish loach (PSYV: 17.0 mg/ml) (You, Zhao, Regenstein, & Ren, ) or blue mussel (FLNEFLHV: 4.95 mg/ml) (Ahn, Kim, & Je, ). However, the EC 50 of HFP was higher than those of antioxidant peptides from protein hydrolysates of grass carp skin (VGGRP: 2.94 mg/ml; GFGPL: 2.25 mg/ml) (Cai et al, ), spotless smoothhound cartilage (GAERP: 3.73 mg/ml; GEREANVM: 1.87 mg/ml; AEVG: 2.30 mg/ml) (Tao, Zhao, Chi, & Wang, ), salmon pectoral fin (TTANIEDRR: 2.50 mg/ml) (Ahn, Cho, & Je, ), and swim bladders of miiuy croaker (FPYLRH: 0.51 mg/ml; GIEWA: 0.78 mg/ml) (Zhao et al, ). HFP could donate electrons or hydrogen radicals for inhibiting the DPPH radical reaction.…”

“…The EC 50 value of HFP was 0.91 mg/ml, which was lower than those of anti‐oxidative peptides from protein hydrolysates of blood clam muscle (QP: 1.41 mg/ml) (Chi, Hu, Wang, Li, & Ding, ), skate cartilage (FIMGPY: 1.43 mg/ml; GPAGDY: 1.33 mg/ml; IVAGPQ: 2.21 mg/ml) (Pan et al, ), and spotless smoothhound muscle (GAA: 1.75 mg/ml; GFVG: 1.31 mg/ml) (Wang et al, ). However, EC 50 value of HFP was higher than those of antioxidant peptides from protein hydrolysates of blood clam muscle (WPP: 0.32 mg/ml) (Chi et al, a), and spotless smoothhound cartilages (GAERP: 0.10 mg/ml; GEREANVM: 0.05 mg/ml; AEVG: 0.07 mg/ml) (Tao et al, ) and muscle (GIISHR: 0.35 mg/ml, ELLI: 0.33 mg/ml; KFPE: 0.46 mg/ml) (Wang et al, ). In consequence, HFP could prevent lipid oxidation via chain breaking reaction.…”

“…2.94 mg/ml; GFGPL: 2.25 mg/ml) (Cai et al, 2015), spotless smoothhound cartilage (GAERP: 3.73 mg/ml; GEREANVM: 1.87 mg/ml; AEVG: 2.30 mg/ml) (Tao, Zhao, Chi, & Wang, 2018), salmon pectoral fin (TTANIEDRR: 2.50 mg/ml) (Ahn, Cho, & Je, 2015), and swim bladders of miiuy croaker (FPYLRH: 0.51 mg/ml; GIEWA: 0.78 mg/ml) . HFP could donate electrons or hydrogen radicals for inhibiting the DPPH radical reaction.…”

“…0.32 mg/ml) (Chi et al, 2015aa), and spotless smoothhound cartilages (GAERP: 0.10 mg/ml; GEREANVM: 0.05 mg/ml; AEVG: 0.07 mg/ml) (Tao et al, 2018) and muscle (GIISHR: 0.35 mg/ml, ELLI: 0.33 mg/ml; KFPE: 0.46 mg/ml) (Wang et al, 2014). In consequence, HFP could prevent lipid oxidation via chain breaking reaction.…”

Identification of antioxidant peptides derived from egg‐white protein and its protective effects on H2O2‐induced cell damage

“…Tao et al . () explored the levels of T‐SOD, CAT, GSH‐Px, and MDA in H 2 O 2 ‐induced HepG2 cells after the treatment with peptides from cartilage protein hydrolysate. The results are in agreement with ours.…”