Purification of lactoperoxidase from bovine milk and investigation of the kinetic properties

Ozdemir H., Aygul I., Kufrevioglu Ö. İ.

PREPARATIVE BIOCHEMISTRY & BIOTECHNOLOGY, cilt.31, sa.2, ss.125-134, 2001 (SCI-Expanded) identifier identifier identifier

  • Yayın Türü: Makale / Tam Makale
  • Cilt numarası: 31 Sayı: 2
  • Basım Tarihi: 2001
  • Doi Numarası: 10.1081/pb-100103378
  • Dergi Adı: PREPARATIVE BIOCHEMISTRY & BIOTECHNOLOGY
  • Derginin Tarandığı İndeksler: Science Citation Index Expanded (SCI-EXPANDED), Scopus
  • Sayfa Sayıları: ss.125-134
  • Atatürk Üniversitesi Adresli: Evet

Özet

Lactoperoxidase (LPO) was purified from bovine milk using Amberlite CG 50 H+ resin, CM Sephadex C-50 ion-exchange chromatography, and Sephadex G-100 gel filtration chromatography. During the purification steps, the activity of enzyme was measured using 2,2 ' -azino-bis (3-ethylbenzthiazoline-6 sulfonic acid) diamonium salt (ABTS) as a chromogenic substrate at pH 6. Optimum pH and optimum temperature values for LPO were determined for ABTS, p-phenylendiamine, catechol, epinephrine, and pyrogallol as substrates, and then K-m and V-max values for the same substrate were obtained by means of Lineweaver-Burk graphics. The purification degree of the enzyme was controlled by SDS-PAGE and R-z (A(412)/A(280)) values.