Zhong-Rui Li

Preparation and evaluation of antioxidant peptides from ethanol-soluble proteins hydrolysate of Sphyrna lewini muscle

To get high yield of ethanol-soluble proteins (EP) and the antioxidant peptides from Sphyrna lewini muscle, orthogonal experiments (L(9)(3)(4)) were applied to optimize the best extraction conditions and enzyme hydrolysis conditions. The yield of EP reached 5.903±0.053% under the optimum conditions of ethanol concentration 90%, solvent to material ratio 20:1, extraction temperature of 40°C and extraction time of 80min. The antioxidant SEPH (EP hydrolysate of S. lewini muscle) was prepared by using papain under the optimum conditions of enzymolysis time 2h, total enzyme dose 1.2%, enzymolysis temperature 50°C and pH 6, and its DPPH radical scavenging activity reached 21.76±0.42% at the concentration of 10mg/ml. Two peptides (F42-3 and F42-5) were isolated from SEPH by using ultrafiltration, anion-exchange chromatography, gel filtration chromatography and RP-HPLC. The structures of F42-3 and F42-5 were identified as Trp-Asp-Arg and Pro-Tyr-Phe-Asn-Lys with molecular weights of 475.50Da and 667.77Da, respectively. F42-3 and F42-5 exhibited good scavenging activity on hydroxyl radical (EC(50) 0.15mg/ml and 0.24mg/ml), ABTS radical (EC(50) 0.34mg/ml and 0.12mg/ml), and superoxide anion radical (EC(50) 0.09mg/ml and 0.11mg/ml), but moderate DPPH radical (EC(50) 3.63mg/ml and 4.11mg/ml). F42-3 and F42-5 were also effectively against lipid peroxidation in the model system and peroxyl free radical scavenging in β-carotene linoleic acid assay. Their high activities were due to the smaller size and the presence of antioxidative amino acids within the peptide sequences.

Purification and characterization of an antioxidant glycoprotein from the hydrolysate of Mustelus griseus

An antioxidant glycoprotein (Fraction AIV-2) with molecular weight of 27.2 kDa was purified from the ethanol-soluble protein hydrolysate of Mustelus griseus muscle. The 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging activity of Fraction AIV-2 reached up to 96.73±2.33% and was higher than that of ascorbic acid at the concentration of 5.0mg/mL. Total protein and carbohydrate contents of Fraction AIV-2 were 62.65±0.63% and 33.49±1.60%, respectively. Seventeen amino acids were identified in Fraction AIV-2, most of which are serine. GC-MS analysis showed that Fraction AIV-2 was composed of fucose, arabinose, galactose, glucose and mannose with the ratio of 1.00:1.53:7.27:9.07:2.09. The FT-IR spectrum of Fraction AIV-2 showed typical characteristics of polysaccharide and protein. For Fraction AIV-2, the changes of ultraviolet absorption curve, amino acid composition after the β-elimination reaction and its deglycosylation with the treatment of N-glycosidase F suggested that both O-glycosidic and N-glycosidic bonds were involved in the polysaccharide and protein moieties.

Characterization of acid-soluble collagens from the cartilages of scalloped hammerhead (Sphyrna lewini), red stingray (Dasyatis akajei), and skate (Raja porosa)

Acid-soluble collagen from the cartilages of scalloped hammerhead (Sphyrna lewini, ASC-S), red stingray (Dasyatis akajei, ASC-D), and skate (Raja porosa, ASC-R) were isolated with yields of 5.64±0.41, 8.72±0.31, and 6.74±0.28% on the basis of wet weight, respectively. ASC-S, ASC-D, and ASC-R had glycine as the major amino acid with contents of 220, 231, and 228 residues/1,000 residues, respectively, and contained imino acid of 174, 155, and 142 residues/1,000 residues, respectively. Three kinds of collagens more likely were comprised of 2 types of collagen (type I and II) and had some differences in primary structures. The denaturation temperatures (Tds) of the ASC-S, ASC-D, and ASC-R were 23.8, 15.1, and 12.1°C, respectively. Maximum solubility of the 3 ASCs in 0.5 M acetic acid was observed at pH 2, and a sharp decrease in solubility above 2%(w/v) was observed in the presence of NaCl. The 3 lyophilized collagens displayed loose, fibrous, and porous structures.

Influence of Different Hydrolysis Processes by Trypsin on the Physicochemical, Antioxidant, and Functional Properties of Collagen Hydrolysates from Sphyrna lewini, Dasyatis akjei, and Raja porosa

ABSTRACT Acid soluble collagen hydrolysates from the cartilages of Sphyrna lewini, Dasyatis akjei, and Raja porosa on different hydrolysis conditions by trypsin were prepared and named as S, D, and R, respectively. The hydrolysates (S1, D1, and R1) obtained from hydrolysis in pH 2.5, 3 h, 37°C presented wonderful foaming and emulsifying capacities, caused by their high average molecular weights (AMWs), high degree of Pro and Lys hydroxylation, and imino acid contents (17.1, 15.3, and 14.1%). At the concentration of 0.1% (w/v), the foaming capacities of S1, D1, and R1 were 104.75 ± 2.57, 63.87 ± 2.73, and 76.87 ± 2.02%; and the emulsifying activity indexes of them were 116.07 ± 1.89, 91.04 ± 1.79, and 123.85 ± 2.14 m2/g, respectively. Conversely, hydrolysates (S2, D2, and R2) obtained from hydrolysis in pH 7.8, 3 min, 37°C exhibited strong scavenging activity on 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical (EC50 0.568, 0.680, and 1.634 mg/mL), hydroxyl radical (EC50 0.253, 0.376, and 0.438 mg/mL), and moderate reducing power (0.0465–0.4702 at the concentration of 5 mg/mL), due to their low AMWs. The results indicated that S1, D1, and R1 could be used as emulsifiers and foaming agents, and S2, D2, and R2 could be used as natural antioxidants in food systems.