Antioxidative properties of peptides obtained from porcine myofibrillar proteins by a protease treatment in an Fe (II)-induced aqueous lipid peroxidation system.

The porcine myofibrillar protein hydrolysate obtained by a papain treatment showed antioxidative activity in a system of linolenic acid peroxidation induced by Fe(2+). The five peptides, DSGVT, IEAEGE, DAQEKLE, EELDNALN, and VPSIDDQEELM, have been characterized as antioxidative peptides (Saiga et al., J. Agric. Food Chem., 51, 3661-3667 (2003)). These peptides were synthesized and their antioxidative activity evaluated. The antioxidative activity of four peptides, excluding DSGVT, was confirmed by their addition at 0.1% to the peroxidation system. To clarify the mechanism for the antioxidative activity of these peptides, their short peptides with amino acid deletions at the C- or N-termini were synthesized. The antioxidative activity gradually decreased with decreasing peptide length. Replacing the charged amino acids in these peptide sequences with Ala also affected their antioxidative activity. We hypothesize that the anions from acidic amino acids and the cations from iron interacted with each other and inactivated the pro-oxidant, Fe (II).

Angiotensin I-converting enzyme-inhibitory peptides obtained from chicken collagen hydrolysate.

In this study, collagen extracted from chicken legs (which are the yellow keratin parts containing a nail) was hydrolyzed with various enzymes, and the angiotensin I-converting enzyme (ACE)-inhibitory activity of each hydrolysate was determined. The hydrolysate by treatment with an Aspergillus species-derived enzyme had the highest activity (IC 50 = 260 microg/mL). The fraction of this hydrolysate obtained by ultrafiltration with a molecular-weight cutoff of 3000 Da (low fraction) had a stronger activity (IC 50 = 130 microg/mL) than the fractionated one. This fraction was further fractionated by HPLC, and the peptides in the fraction with high ACE-inhibitory activity were identified. The amino acid sequences of the four peptides were identified using a protein sequencer. These peptides were synthesized to confirm their ACE-inhibitory activities; this showed that peptides with a Gly-Ala-Hyp-Gly-Leu-Hyp-Gly-Pro sequence had the highest activity (IC 50 = 29 microM). When the low fraction was administered to spontaneous hypertensive rats, a decrease in their blood pressure was observed after 2 h of administration, and a significant decrease in blood pressure (-50 mmHg) was observed after 6 h. Moreover, long-term administration studies indicated that the low fraction showed a significant suppression of increased blood pressure.

Royal jelly supplementation improves lipoprotein metabolism in humans.

Royal jelly (RJ) has several physiological effects and is widely used in commercial medical products and health foods. We examined the effects of RJ supplementation on serum lipoprotein metabolism in humans. Fifteen volunteers were divided into an RJ intake group (n=7) and a control group (n=8). The RJ group took 6 g per day for 4 wk. Their serum total cholesterol (TC) and serum low-density lipoprotein (LDL) decreased significantly compared with those of the control group (p<0.05). There were no significant differences in serum high-density lipoprotein (HDL) or triglyceride concentrations. Moreover, the relationship between the serum cholesterol and lipoprotein levels was investigated. Among the lipoprotein fractions, small very-low-density lipoprotein was decreased (p<0.05) after RJ intake. Our results suggest that dietary RJ decreases TC and LDL by lowering small VLDL levels.

Action mechanism of an angiotensin I-converting enzyme inhibitory peptide derived from chicken breast muscle.

In a previous study, we isolated the inhibitory peptide (P4 peptide, Gly-Phe-Hyp-Gly-Thr-Hyp-Gly-Leu-Hyp-Gly-Phe) for angiotensin I-converting enzyme (ACE) from chicken breast muscle extract possessing hypotensive activity for spontaneously hypertensive rats (SHRs). This study was performed to elucidate the peptide's action mechanisms of inhibiting ACE. Intravenous administration of synthetic P4 peptide resulted in significant drops in the blood pressures of SHRs. As Dixon plots indicate, the P4 peptide showed high affinity toward ACE (K(i) = 11.48 microM) and only 10% of the total amount of the P4 peptide was decomposed. The analyses of the relationship between the ACE inhibitory activity and structure of the P4 peptide clarified that Hyp-Gly-Leu-Hyp-Gly-Phe showed a stronger activity (IC50 = 10 microM) than the P4 peptide (IC50 = 46 microM). When Phe at the C-terminus of the P4 peptide was deleted, IC50 changed to 25000 microM, indicating that Phe at the C-terminus of the peptide is very important for ACE inhibitory activity.

Antioxidant activity of peptides obtained from porcine myofibrillar proteins by protease treatment.

Hydrolysates obtained from porcine myofibrillar proteins by protease treatment (papain or actinase E) exhibited high antioxidant activity in a linolenic acid peroxidation system induced by Fe(2+). Hydrolysates produced by both papain and actinase E showed higher activities at pH 7.1 than at pH 5.4. The antioxidant activity of the papain hydrolysate was almost the same as that of vitamin E at pH 7.0. These hydrolysates possessed 1,1-diphenyl-2-picrylhydrazyl radical scavenging activity and chelating activity toward metal ions. Antioxidant peptides were separated from the papain hydrolysate by ion exchange chromatography. The acidic fraction obtained by this method exhibited higher activity than the neutral or basic fractions. Antioxidant peptides in the acidic fraction were isolated by high-performance liquid chromatography on an ODS column and shown to possess the structures DSGVT, IEAEGE, DAQEKLE, EELDNALN, and VPSIDDQEELM. The DAQEKLE peptide showed the highest activity among these peptides.

Angiotensin I-converting enzyme inhibitory peptides in a hydrolyzed chicken breast muscle extract.

The blood pressure of spontaneously hypertensive rats (SHRs) decreased after oral administration of an extract prepared from chicken breast muscle, falling maximally to 50 mmHg lower than before. This effect continued for at least 4 h after administration. The peptides possessing hypotensive activity in the chicken extract were examined by measuring the inhibitory activity (IC(50)) against angiotensin I-converting enzyme (ACE). The inhibitory activity of the chicken extract was 1060 mg%, whereas the activity of the extract treated with an Aspergillus protease and gastric proteases (trypsin, chymotrypsin, and intestinal juice) became stronger, reaching 1.1 mg%. Peptides in this hydrolysate of the extract were isolated by HPLC on a reversed-phase column, and their N-terminal sequences were analyzed. Three peptides possessed a common sequence, Gly-X-X-Gly-X-X-Gly-X-X, which was homologous with that of collagen. The peptide Gly-Phe-Hyp-Gly-Thr-Hyp-Gly-Leu-Hyp-Gly-Phe showed the strongest inhibitory activity (IC(50) = 42 microM).