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Journals / Turkish Journal of Chemistry / 2015 / Cilt: 39 - Sayı: 1

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
Pages
149–158
Publication Date
2015-03-15
DOI
—
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AI Summary & Key Takeaways

AI Insight
⚡ TL;DR(Too Long; Didn't Read)

SDS-PAGE results showed a single band and the molecular weight of NADPH-cytochrome P450 reductase was 70 kDa.

📌 3 Key Takeaways

🎯
Objective & Hypothesis: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.
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Key Findings & Data:SDS-PAGE results showed a single band and the molecular weight of NADPH-cytochrome P450 reductase was 70 kDa.
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Conclusion & Impact:IC50 values and Ki values of metals showing an inhibitory effect were calculated.
🔬Design:Research Article
📍Scope:Comprehensive Sample
⚙️Variables:NADPH-cytochrome P450 reductase · Lake Van fish · inhibition

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.

Özet

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.

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Cite This Article

KUZU, M., ÇİFTCİ, M. (2015). 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. Turkish Journal of Chemistry, Cilt: 39 - Sayı: 1, 149–158. https://ajindex.com/articles/184742

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