title: Improved chromatographic method for purification of lactoperoxidase from different milk sources.
abstract: Our previous studies showed that sulfanilamide is a new competitive inhibitor of and can be used in the purification of lactoperoxidase (LPO, EC1.11.1.7) from milk. However, this method has some disadvantages like a lower purification factor. The aim of the present study is to improve the purification process of milk LPO from different sources. For this purpose, 16 commercial sulfanilamide derivatives were selected for inhibition studies to determine the best inhibitor of bovine LPO by calculating kinetic parameters. A cyanogen bromide-activated Sepharose 4B affinity matrix was synthesized by coupling with each competitive inhibitor. Among the inhibitors, 5-amino-2-methylbenzenesulfonamide and 2-chloro-4-sulfamoylaniline were used as ligands for the purification of LPO from bovine, buffalo, cow, and goat milks with 1059.37, 509.09, 232.55, and 161.90, and 453.12-, 151.86-, 869.00-, and 447.57-fold, respectively. Our results show that 5-amino-2-methylbenzenesulfonamide, 2-chloro-4-sulfamoylaniline, and 5-amino-1-naphthalenesulfonamide are the best inhibitors for one-step purification of the enzyme.
authors.affiliation: a Department of Chemistry, Faculty of Sciences , Atatürk University , Erzurum , Turkey.|b Department of Chemistry, Faculty of Sciences , Istanbul Medeniyet University , Istanbul , Turkey.
authors.forename: Zeynep
authors.initials: Z
authors.lastname: Köksal
authors.affiliation: c Department of Bioengineering, Faculty of Engineering and Architecture , Sinop University , Sinop , Turkey.
authors.forename: Hande
authors.initials: H
authors.lastname: Usanmaz
authors.affiliation: a Department of Chemistry, Faculty of Sciences , Atatürk University , Erzurum , Turkey.
authors.forename: Songül
authors.initials: S
authors.lastname: Bayrak
authors.affiliation: a Department of Chemistry, Faculty of Sciences , Atatürk University , Erzurum , Turkey.
authors.forename: Hasan
authors.initials: H
authors.lastname: Ozdemir
mesh_terms: D000818:Animals; D002853:Chromatography, Liquid / Q000379:methods*; D004591:Electrophoresis, Polyacrylamide Gel; D007700:Kinetics; D007784:Lactoperoxidase / Q000037:antagonists & inhibitors / Q000737:chemistry / Q000302:isolation & purification*; D008892:Milk / Q000201:enzymology*; D008970:Molecular Weight
chemical_list: D007784:Lactoperoxidase
keywords: Affinity chromatography; enzyme purification; lactoperoxidase; mammalian milk