Purification and characterization of NADPH-cytochrome P450 reductase from Lake Van fish liver microsomes and investigation of some chemical and metals' effects on the enzyme activity

Müslüm KUZU; Mehmet ÇİFTCİ

Article Details

Purification and characterization of NADPH-cytochrome P450 reductase from Lake Van fish liver microsomes and investigation of some chemical and metals' effects on the enzyme activity

Authors:

Müslüm KUZU | Mehmet ÇİFTCİ

Publisher:

TUBITAK

Journal:

Turkish Journal of Chemistry 2015 Vol.39 Issue 1, pp.149-158

Document Type:

Article

Publication Date:

3/2015

Document Language:

Turkish

Document ID (Org No.):

2000726985

Frequency:

Bimonthly

eISSN:

1303-6130

pISSN:

1300-0527

Abstract:

 

NADPH-cytochrome P450 reductase was purified from Lake Van fish liver  microsomes by primary and secondary DEAE-cellulose column chromatograph with  20.46 m M/min/mg enzyme specific activities, 54.4% purification  yield, and purification of 38-fold. The purity of the enzyme was  established, and its monomer molecular weight was determined by  SDS-polyacrylamide gel electrophoresis. SDS-PAGE results showed a single  band and the molecular weight of NADPH-cytochrome P450 reductase was 70 kDa.  In addition, optimum ionic strength, optimum pH, optimum temperature, and  stable pH values were determined for the enzyme in the kinetic studies  performed. KM and Vmax were determined for NADPH and cytochrome c.  Effects of some metals ions, antibiotics, and some other drugs used in  aquarium fisheries on the activity of the enzyme were investigated. IC50  values and Ki values of metals showing an inhibitory effect were  calculated.

 

NADPH-cytochrome P450 reductase was purified from Lake Van fish liver  microsomes by primary and secondary DEAE-cellulose column chromatograph with  20.46 m M/min/mg enzyme specific activities, 54.4% purification  yield, and purification of 38-fold. The purity of the enzyme was  established, and its monomer molecular weight was determined by  SDS-polyacrylamide gel electrophoresis. SDS-PAGE results showed a single  band and the molecular weight of NADPH-cytochrome P450 reductase was 70 kDa.  In addition, optimum ionic strength, optimum pH, optimum temperature, and  stable pH values were determined for the enzyme in the kinetic studies  performed. KM and Vmax were determined for NADPH and cytochrome c.  Effects of some metals ions, antibiotics, and some other drugs used in  aquarium fisheries on the activity of the enzyme were investigated. IC50  values and Ki values of metals showing an inhibitory effect were  calculated.

Article Details

Purification and characterization of NADPH-cytochrome P450 reductase from Lake Van fish liver microsomes and investigation of some chemical and metals' effects on the enzyme activity

Authors: Müslüm KUZU | Mehmet ÇİFTCİ

Publisher: TUBITAK

Journal: Turkish Journal of Chemistry 2015 Vol.39 Issue 1, pp.149-158

Document Type: Article

Publication Date: 3/2015

Document Language: Turkish

Document ID (Org No.): 2000726985

Frequency: Bimonthly

eISSN: 1303-6130

pISSN: 1300-0527

Issues

Authors:

Müslüm KUZU | Mehmet ÇİFTCİ

Publisher:

TUBITAK

Journal:

Turkish Journal of Chemistry 2015 Vol.39 Issue 1, pp.149-158

Document Type:

Article

Publication Date:

3/2015

Document Language:

Turkish

Document ID (Org No.):

2000726985

Frequency:

Bimonthly

eISSN:

1303-6130

pISSN:

1300-0527