Improved chromatographic method for purification of lactoperoxidase from different milk sources

Koksal Z., Usanmaz H., BAYRAK S., ÖZDEMİR H.

PREPARATIVE BIOCHEMISTRY & BIOTECHNOLOGY, cilt.47, sa.2, ss.129-136, 2017 (SCI-Expanded) identifier identifier identifier

  • Yayın Türü: Makale / Tam Makale
  • Cilt numarası: 47 Sayı: 2
  • Basım Tarihi: 2017
  • Doi Numarası: 10.1080/10826068.2016.1185732
  • Dergi Adı: PREPARATIVE BIOCHEMISTRY & BIOTECHNOLOGY
  • Derginin Tarandığı İndeksler: Science Citation Index Expanded (SCI-EXPANDED), Scopus
  • Sayfa Sayıları: ss.129-136
  • Anahtar Kelimeler: Affinity chromatography, enzyme purification, lactoperoxidase, mammalian milk, AFFINITY-CHROMATOGRAPHY, BOVINE LACTOPEROXIDASE, STEP PURIFICATION, PEROXIDASE, SYSTEM
  • Atatürk Üniversitesi Adresli: Evet

Özet

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-4sulfamoylaniline, and 5-amino-1-naphthalenesulfonamide are the best inhibitors for one-step purification of the enzyme.