Stabilization of horseradish peroxidase by covalent conjugation with dextran aldehyde against temperature and pH changes

Altikatoglu M., Arioz C., Basaran Y., Kuzu H.

CENTRAL EUROPEAN JOURNAL OF CHEMISTRY, cilt.7, sa.3, ss.423-428, 2009 (SCI-Expanded) identifier identifier

  • Yayın Türü: Makale / Tam Makale
  • Cilt numarası: 7 Sayı: 3
  • Basım Tarihi: 2009
  • Doi Numarası: 10.2478/s11532-009-0041-z
  • Dergi Adı: CENTRAL EUROPEAN JOURNAL OF CHEMISTRY
  • Derginin Tarandığı İndeksler: Science Citation Index Expanded (SCI-EXPANDED), Scopus
  • Sayfa Sayıları: ss.423-428
  • Yıldız Teknik Üniversitesi Adresli: Evet

Özet

Stabilization of Horseradish Peroxidase (HRP; EC 1.11.1.7) against temperature and pH via the formation of the conjugates obtained by multipoint covalent bonding of dextran aldehyde (DA) to the enzyme were studied. Hence, three different molar weighted dextrans (17.5 kD, 75 kD, 188 Q were covalently bonded to purified enzyme with different molar ratios (n(HRP)/n(DA) 20/1, 10/1, 1/1, 1/5, 1/10, 1/15, 1/20). The thermal stabilities of the obtained conjugates were evaluated with the activities determined at different temperatures (25, 30, 35, 40, 50, 60, 70, 80 degrees C) applying 60 minutes incubation time. Conjugates formed were characterized by gel-permeation chromatography (GPC) and fluorescence techniques. The conjugate synthesized using dextran 75 kDa with n(HRP)/n(DA) 1/10 molar ratio showed better thermal stability than other conjugates and purified enzyme at pH 7. This conjugate also has wider activity pH range than purified enzyme. In addition, mentioned conjugate at pH 7 had very long storage lifetime compared to purified enzyme at +4 degrees C and room temperature; which is considered a favorable feature for usage in practice.