Megías Cristina, Pedroche Justo, Yust María del Mar, Alaiz Manuel, Girón-Calle Julio, Millan Francisco, Vioque Javier
Instituto de la Grasa (CSIC), Padre García Tejero 4, 41012-Sevilla, Spain.
J Agric Food Chem. 2006 Sep 20;54(19):7120-4. doi: 10.1021/jf061488b.
A lung extract rich in angiotensin converting enzyme (ACE) and pure ACE were immobilized by reaction with the activated support 4 BCL glyoxyl-agarose. These immobilized ACE derivatives were used for purification of ACE inhibitory peptides by affinity chromatography. The immobilized lung extract was used to purify inhibitory peptides from sunflower and rapeseed protein hydrolysates that had been obtained by treatment of protein isolates with alcalase. The ACE binding peptides that were retained by the derivatives were specifically released by treatment with the ACE inhibitor captopril and further purified by reverse-phase C18 HPLC chromatography. Inhibitory peptides with IC50 50 and 150 times lower than those of the original sunflower and rapeseed hydrolysates, respectively, were obtained. The derivative prepared using pure ACE was used for purification of ACE inhibitory peptides from the same type of sunflower protein hydrolysate. ACE binding peptides were released from the ACE-agarose derivatives by treatment with 1 M NaCl and had an IC50 a little higher than those obtained using immobilized extract and elution with captopril. Affinity chromatography facilitated the purification of ACE inhibitory peptides and potentially other bioactive peptides present in food proteins.
富含血管紧张素转换酶（ACE）的肺提取物和纯ACE通过与活化载体4 - BCL乙醛酸琼脂糖反应进行固定化。这些固定化的ACE衍生物用于通过亲和色谱法纯化ACE抑制肽。固定化的肺提取物用于从向日葵和菜籽蛋白水解物中纯化抑制肽，这些水解物是通过用碱性蛋白酶处理蛋白质分离物而获得的。被衍生物保留的ACE结合肽通过用ACE抑制剂卡托普利处理而特异性释放，并通过反相C18高效液相色谱法进一步纯化。分别获得了IC50比原始向日葵和菜籽水解物低50倍和150倍的抑制肽。使用纯ACE制备的衍生物用于从相同类型的向日葵蛋白水解物中纯化ACE抑制肽。通过用1 M NaCl处理从ACE - 琼脂糖衍生物中释放ACE结合肽，其IC50略高于使用固定化提取物并用卡托普利洗脱所获得的IC50。亲和色谱法有助于纯化ACE抑制肽以及食品蛋白质中潜在的其他生物活性肽。
