- 期刊
去脂蛋黃蛋白質酵素水解物抗氧化、抑制ACE及其胺基酸序列
Evaluation the Functional Properties of Delipid Egg Yolk Protein Enzymatic Hydrolysate on Antioxidant Ability and Inhibition of Angiotensin I-Converting Enzyme
摘要
去脂蛋黃蛋白質(Delipid egg yolk protein, DEYP)，分別以Alcalase 或Papain水解，依基質溫度1.5%、酵素基質比(E/S，1/1000) 和pH 分別為9.0及6.0，於60°C進行酵素水解1~8小時。比較抗氧化與抑制ACE活性的能力，結果顯示以Alcalase及Papain水解1~8小時，其水解物清除DPPH能力及還原力均不佳，然具良好的螯合亞鐵離子能力，其IC_(50)分別為80~129及26~55μg/mL，抑制ACE活性之IC_(50)則分別為0.40~2.01及0.42~1.21mg/mL。再將這兩種水解物經分子量5.0KDa 超過濾膜區分，測得分子量小於5.0KDa 之胜肽，其抑制ACE 活性之IC_(50)分別為0.045及0.039 mg/mL;最後再將分子量小於5.0 KDa 之胜肽經HPLC區分純化分別收集四個區段，以nano LC/ESI/MS進行胜肽定序，測得第一區段Lys-Pro-Gln-Met-Thr-Glu-Glu-Gln，Lys-Pro-Gln-Met-Thr-Glu- Glu-Gln-IIe-Lys，Ala-Val-Asn-Lys-Val-Ser-Pro-Arg-Ala-Gly，Glu-Ala-Val-Asn-Lys-Val-Ser-Pro-Arg-Ala-Gly。第二區段Ala-Pro-Ala-Glu-Glu-Arg-Glu-Val-Gly，Lys-Pro-Gln-Met-Thr-Glu-Glu-Gln-lIe-Lys。第三區段Ala-Ala-Glu-Gln-Ala-Glu-Gln-lle-Asp-Tyr-Gln-Arg，Tyr-Pro-Glu-Lys-Asp-Glu-Pro-Lcu-Asp。第四區段Lcu-Pro-Val-Gly-Pro-Arg-Ilc-Pro-Ala-Scr，Lcu-Pro-Val-Gly-Pro-Arg-Ile-Pro-Ala-Ser-Glu等小胜肽。
並列摘要
To investigate the functional properties of delipid egg yolk protein (DEYP), DEYP were hydrolyzed with alcalase or papain using the optimal hydrolysis parameter below: substrate concentration 1.5 %, enzyme to substrate (E/S) ratio 1/1000, pH 9.0 (alcalse) and pH 6.0 (papain), at 60°C for 1~8 hours. The anti-oxidative capacity and angiotensin I-converting enzyme (ACE) inhibitory activity of the hydrolysates were then evaluated. The results showed that the DPPH scavenging activity and reducing power of 1~8 hours alcalase or papain hydrolysates were poor. On the other hand, their ferrous ion chelating abilities were good, with IC_(50) of 80~129μg/mL and 26~55μg/mL, respectively. Moreover, the ACE inhibitory ability were between IC_(50) of 0.40~2.01 mg/mL and 0.42~1.21 mg/mL. The hydrolysates from alcalse or papain were further separated using 5 kDa ultrafiltration membrane, and the peptides with molecular weight less than 5.0 kDa were collected and evaluated. The ACE inhibitory abilities of these peptides were at IC_(50) of 0.045 mg/mL and 0.039 mg/mL. Finally, these small peptides (＜5.0 kDa) were purified using HPLC and 4 fractions were collected, followed by sequencing using nano LC/ESI/MS. The peptide sequences are: [Fraction 1] Lys-Pro-Gln-Met-Thr-Glu-Glu-Gin, Lys-Pro-Gln-Met-Thr-Glu-Glu-Gln-Ile-Lys, Ala-Val-Asn-Lys-Val-Ser-Pro-Arg-Ala-Gly, Glu-Ala-Val-Asn-Lys-Val-Ser-Pro-Arg-Ala-Gly. [Fraction 2] Ala-Pro-Ala-Glu-Glu-Arg-GluVal- Gly, Lys-Pro-Gln-Met-Thr-Glu-Glu-Gln-Ile-Lys. [Fraction 3] Ala-Ala-Glu-Gln-Ala-GluGln-Ile- Asp-Tyr-Gln-Arg, Tyr-Pro-Glu-Lys-Asp-Glu-Pro-Leu-Asp. [Fraction 4] Leu-Pro-Val-Gly-Pro-Arg-lle-Pro-Ala-Ser and Leu-Pro-Val-Gly-Pro-Arg-Ile-Pro-Ala-Ser-Glu.