Affinity purification of angiotensin converting enzyme inhibitory peptides using immobilized ACE.
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.
Megías
Cristina
C
Instituto de la Grasa (CSIC), Padre García Tejero 4, 41012-Sevilla, Spain.
Pedroche
Justo
J
Yust
María del Mar
Mdel M
Alaiz
Manuel
M
Girón-Calle
Julio
J
Millan
Francisco
F
Vioque
Javier
J
D000806:Angiotensin-Converting Enzyme Inhibitors / Q000302:isolation & purification*; D000818:Animals; D029741:Brassica rapa / Q000737:chemistry; D002846:Chromatography, Affinity / Q000379:methods*; D004800:Enzymes, Immobilized; D006368:Helianthus / Q000737:chemistry; D008168:Lung / Q000201:enzymology; D010455:Peptides / Q000302:isolation & purification*; D007703:Peptidyl-Dipeptidase A; D011817:Rabbits; D012639:Seeds / Q000737:chemistry; D013552:Swine
D000806:Angiotensin-Converting Enzyme Inhibitors; D004800:Enzymes, Immobilized; D010455:Peptides; D007703:Peptidyl-Dipeptidase A