酪蛋白ACE抑制肽的分离纯化.pdfVIP

中国科技论文在线 酪蛋白 ACE 抑制肽的分离纯化# 洪旭，卢蓉蓉** （江南大学食品学院，江苏 无锡 214122） 5 摘要：目的：对酪蛋白酶解物进行分离纯化得到新的具有明确氨基酸序列的高活性ACE 抑 制短肽。方法：在 Alcalase 2.4L 与 Neutrase 1.5MG 蛋白酶分两步酶解酪蛋白制备 ACE 抑制 肽的基础上，依次采用大孔吸附树脂、凝胶层析、制备和分析型反相高效液相色谱 （RP-HPLC ）分离纯化，并利用液相- 电喷雾电离-质谱联用（LC-ESI/MS/MS ）对分离得到 的组分进行质谱分析，获得ACE 抑制肽的氨基酸序列。结果：经分离得到 4 条酪蛋白 ACE 10 抑制短肽（D3a 、D3b、D5a 、D5b ），其氨基酸序列分别为LY 、WKQ 、WQV 和 LQSW ， ACE 抑制活性 IC50 值分别为 23.8 、48.5、40.8 和 28.0 μg/mL。结论：经本次分离纯化得到 的肽均具有较小的分子量和较高的 ACE 抑制活性，且 ACE 抑制肽 WKQ 、WQV 和 LQSW 均为首次报道。 关键词：酪蛋白；ACE 抑制肽；分离纯化；氨基酸序列 15 中图分类号：TS252.9 Isolation and Purification of ACE Inhibitory Peptides from Casein Hydrolysate HONG Xu, LU Rongrong 20 (School of Food Science and Technology, Jiangnan University, JiangSu WuXi 214122) Abstract: Object: For separation and purification of casein hydrolysate to obtain the novel amino acid sequences of ACE inhibitory peptides with high-activity and small molecular weight. Method: On the basis of the ACE inhibitory peptides from casein hydrolysed by Alcalase 2.4L and Neutrase 1.5MG proteases, the casein hydrolysate were purified by macroporous adsorption resin, 25 sephadex G-15 gel filtration column, preparative and analytic RP-HPLC. The amino acid sequences of ACE inhibitory peptides were identified by LC-ESI/MS/MS. Results: Four casein ACE inhibitory peptides were obtained with amino acid sequences LY, WKQ, WQV and LQSW, the IC50 of ACE inhibiting activity were 23.8 μg/mL, 48.5 μg/mL, 40.8 μg/mL and 28.0 μg