Purification and Partial Characterization of Catalase from Chicken Erythrocytes and the Effect of Various Inhibitors on Enzyme Activity

Tülin AYDEMİR; Kevser KURU

Makale detayları

Purification and Partial Characterization of Catalase from Chicken Erythrocytes and the Effect of Various Inhibitors on Enzyme Activity

Yazarlar:

Tülin AYDEMİR | Kevser KURU

Yayıncı:

TUBITAK

Dergi:

Turkish Journal of Chemistry 2003 Vol.27 Issue 1, pp.85-98

Belge Tipi:

Makale

Yayın Tarihi:

12/2002

Belge Dili:

Türkçe

Document ID (Org No.):

2000729685

Sıklık:

İki Aylık

eISSN:

1303-6130

pISSN:

1300-0527

Özet:

 

Catalase plays a major role in the protection of tissues  from the toxic effects of H2O2 and partially reduced oxygen  species.  A nearly 136-fold enzyme purification was obtained from  chicken erythrocyte by acetone precipitation, ethanol-chloroform  treatment, CM-cellulose and Sephadex G-200 chromatography.  The  specific activity of purified enzyme was 42,556 U/mg.  The molecular  weight of the native chicken erythrocyte catalase was estimated at 240  kDa by gel filtration.  SDS-gel electrophoresis results indicated that  chicken erythrocyte catalase consists of four apparently identical  subunits, with a molecular weight of around 57.5 kDa.  The optical  spectrum of the purified enzyme shows a Soret band at 406 nm, which is  the characteristic for the heme group.  Dithionite treatment of the  enzyme resulted in the reduction of enzyme.  The Km of chicken  erythrocyte catalase was 33 mM H2O2.  The maximal activity of  catalase was observed between pH 6.0 and 8.0.  Enzyme activity was  stable at temperatures between 10 and 30°C. The activity of purified  catalase was inhibited by azide, cyanide, b -mercaptoethanol,  dithiotreitol (DTT) and iodoacetamide.

 

Catalase plays a major role in the protection of tissues  from the toxic effects of H2O2 and partially reduced oxygen  species.  A nearly 136-fold enzyme purification was obtained from  chicken erythrocyte by acetone precipitation, ethanol-chloroform  treatment, CM-cellulose and Sephadex G-200 chromatography.  The  specific activity of purified enzyme was 42,556 U/mg.  The molecular  weight of the native chicken erythrocyte catalase was estimated at 240  kDa by gel filtration.  SDS-gel electrophoresis results indicated that  chicken erythrocyte catalase consists of four apparently identical  subunits, with a molecular weight of around 57.5 kDa.  The optical  spectrum of the purified enzyme shows a Soret band at 406 nm, which is  the characteristic for the heme group.  Dithionite treatment of the  enzyme resulted in the reduction of enzyme.  The Km of chicken  erythrocyte catalase was 33 mM H2O2.  The maximal activity of  catalase was observed between pH 6.0 and 8.0.  Enzyme activity was  stable at temperatures between 10 and 30°C. The activity of purified  catalase was inhibited by azide, cyanide, b -mercaptoethanol,  dithiotreitol (DTT) and iodoacetamide.

Makale detayları

Purification and Partial Characterization of Catalase from Chicken Erythrocytes and the Effect of Various Inhibitors on Enzyme Activity

Yazarlar: Tülin AYDEMİR | Kevser KURU

Yayıncı: TUBITAK

Dergi: Turkish Journal of Chemistry 2003 Vol.27 Issue 1, pp.85-98

Belge Tipi: Makale

Yayın Tarihi: 12/2002

Belge Dili: Türkçe

Document ID (Org No.): 2000729685

Sıklık: İki Aylık

eISSN: 1303-6130

pISSN: 1300-0527

Sayı

Yazarlar:

Tülin AYDEMİR | Kevser KURU

Yayıncı:

TUBITAK

Dergi:

Turkish Journal of Chemistry 2003 Vol.27 Issue 1, pp.85-98

Belge Tipi:

Makale

Yayın Tarihi:

12/2002

Belge Dili:

Türkçe

Document ID (Org No.):

2000729685

Sıklık:

İki Aylık

eISSN:

1303-6130

pISSN:

1300-0527