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Inhibitory Effects of Ofloxacin and Cefepime on Enzyme Activity of 6-Phosphogluconate Dehydrogenase from Chicken Liver

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In this study, effects of some antibiotics, namely, ofloxacin, cefepime, cefazolin, and ampicillin on the in vitro enzyme activity of 6-phosphogluconate dehydrogenase have been investigated. For this purpose, 6-phosphogluconate dehydrogenase was purified from chicken liver 535-fold with a yield of 18% by using ammonium sulphate precipitation, 2',5'-ADP Sepharose 4B affinity chromatography, and Sephadex G-200 gel filtration chromatography. In order to check the purity of the enzyme, SDS polyacylamide gel electrophoresis (SDS-PAGE) was performed. This analysis revealed a highly pure enzyme band on the gel. Among the antibiotics, ofloxacin and cefepime exhibited inhibitory effects, but cefazolin and ampicillin showed neither important inhibitory nor activatory effects on the enzyme activity. The measured I50 values by plotting activity percent vs. inhibitor concentration, [I50] were 0.1713 mM for ofloxacin and 6.0028 mM for cefepime. Inhibition constants, Ki, for ofloxacin and cefepime were also calculated as 0.2740 ± 0.1080 mM and 12.869 ± 16.6540 mM by means of Lineweaver-Burk graphs, and inhibition types of the antibiotics were found out to be noncompetitive and competitive, respectively. It has been understood from the calculated inhibitory parameters that the purified chicken enzyme has been quite inhibited by these two antimicrobials.

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Keywords: 6-phosphogluconate dehydrogenase; cefepime; chicken; liver; ofloxacin

Document Type: Research Article

Affiliations: Atatürk University, Biotechnology Application and Research Center, 25240-Erzurum, Turkey.

Publication date: February 1, 2007

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  • Protein & Peptide Letters publishes short papers in all important aspects of protein and peptide research, including structural studies, recombinant expression, function, synthesis, enzymology, immunology, molecular modeling, drug design etc. Manuscripts must have a significant element of novelty, timeliness and urgency that merit rapid publication. Reports of crystallisation, and preliminary structure determinations of biologically important proteins are acceptable. Purely theoretical papers are also acceptable provided they provide new insight into the principles of protein/peptide structure and function.
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