title: Affinity purification of copper chelating peptides from chickpea protein hydrolysates.
abstract: Chickpea protein hydrolysates obtained with alcalase and flavourzyme were used for purification of copper chelating peptides by affinity chromatography using copper immobilized on solid supports. The chelating activity of purified peptides was indirectly measured by the inhibition of beta-carotene oxidation in the presence of copper. Two protein hydrolysates, obtained after 10 and 100 min of hydrolysis, were the most inhibitory of beta-carotene oxidation. Purified copper chelating peptides from these protein hydrolysates contained 19.7 and 35.1% histidine, respectively, in comparison to 2.7 and 2.6% in the protein hydrolysates. Chelating peptides from hydrolysate obtained after 10 min of hydrolysis were the most antioxidative being 8.3 times more antioxidative than the hydrolysate, while chelating peptides purified from protein hydrolysate obtained after 100 min were 3.1 times more antioxidative than its hydrolysate. However, the histidine content was higher in peptides derived from the 100 min hydrolysate (19.7 against 35.1% in 10 min hydrolysate), indicating that this amino acid is not the only factor involved in the antioxidative activity, and other factors such as peptide size or amino acid sequence are also determinant. This manuscript shows that affinity chromatography is a useful procedure for purification of copper chelating peptides. This method can be extended to other metals of interest in nutrition, such as calcium, iron, or zinc. Purified chelating peptides, in addition to their antioxidative properties, may also be useful in food mineral fortification for increasing the bioavailability of these metals.
authors.affiliation: Instituto de la Grasa CSIC, Avda Padre García Tejero 4, 41012-Sevilla, Spain.
authors.forename: Cristina
authors.initials: C
authors.lastname: Megías
authors.forename: Justo
authors.initials: J
authors.lastname: Pedroche
authors.forename: Maria M
authors.initials: MM
authors.lastname: Yust
authors.forename: Julio
authors.initials: J
authors.lastname: Girón-Calle
authors.forename: Manuel
authors.initials: M
authors.lastname: Alaiz
authors.forename: Francisco
authors.initials: F
authors.lastname: Millan
authors.forename: Javier
authors.initials: J
authors.lastname: Vioque
mesh_terms: D000975:Antioxidants / Q000494:pharmacology; D002614:Chelating Agents / Q000302:isolation & purification* / Q000494:pharmacology; D002846:Chromatography, Affinity; D029863:Cicer / Q000737:chemistry*; D003300:Copper; D010455:Peptides / Q000302:isolation & purification* / Q000494:pharmacology; D010940:Plant Proteins / Q000737:chemistry*; D011492:Protein Hydrolysates / Q000737:chemistry / Q000494:pharmacology; D013381:Subtilisins / Q000378:metabolism
chemical_list: D000975:Antioxidants; D002614:Chelating Agents; D010455:Peptides; D010940:Plant Proteins; D011492:Protein Hydrolysates; D003300:Copper; D013381:Subtilisins