酪蛋白双酶水解物ACE 抑制肽的分离纯化.pdfVIP

50 2012, Vol. 33, No. 09 食品科学 ※基础研究 ACE 杨 铭，胡志和 * (天津市食品生物技术重点实验室，天津商业大学生物技术与食品科学学院，天津 300134) 采用胃蛋白酶和胰蛋白酶依次对酪蛋白进行双酶水解，制备 ACE 抑制肽。水解物经截留分子质量 6ku 的 超滤膜初步分离，再通过 Sephadex G-15 进一步纯化，体外测定各分离产物 ACE 活性的半数抑制质量浓度(IC50 值) 。 纯化得到的各组分经毛细管电泳分析肽谱、Q-TOF LC/MS 检测分子质量范围。结果显示：双酶水解产物 IC50 值 为560 μg/mL ，超滤流出物IC50 值为250 μg/mL；Sephadex G-15 分离得到3 个组分，组分Ⅰ的IC50 值为 123.41μg/mL ， 含有 19 个肽段；组分Ⅱ的 IC50 值为 66.67 μg/mL ，含有 14 个肽段；组分Ⅲ的 IC50 值为 64.29 μg/mL ，含有5 个肽段。 Q-TOF LC/MS 测得纯化组分的分子质量范围为 400～800u 。 酪蛋白；酪蛋白水解物；A C E 抑制肽；胃蛋白酶；胰蛋白酶 Separation and Purification of ACE Inhibitory Peptides from Dual-Enzymatic Casein Hydrolysate YANG Ming ，HU Zhi-he* (Tianjin Key Laboratory of Food and Biotechnology, College of Biotechnology and Food Science, Tianjin University of Commerce, Tianjin 300134, China) Abstract ：ACE inhibitory peptides were prepared from casein by sequential hydrolysis with pepsin and trypsin. Casein hydrolysate was preliminarily separated with 6 ku MWCO ultrafiltration membrane and further purified by Sephadex G-15 column chromatography. The ACE inhibitory activity (IC50) of the separated fractions was determined by an in vitro assay, and their peptide profiles and molecular weights were measured by capillary electrophoresis (CE) and Q-TOF LC/MS, respectively. The results showed that IC50 values of casein hydrolysate, filtrate, and three fractions obtained by Sephadex G-15 column chromatography, factions Ⅰ, Ⅱ and Ⅲ were 560, 250, 123.41, 66.67 μg/mL and 64.29 μg/mL, respectively. Factions Ⅰ, Ⅱ and Ⅲ were composed of 19, 14 and 5 peptides, respectively and their molecular weights ranged from 400 to 800 u. Key words ：casein ；casein hydrolysate ；ACE inhibitory peptide ；p