Purification and characterization of glucose 6-phosphate dehydrogenase enzyme from rainbow trout (Oncorhynchus mykiss) liver and investigation of the effects of some metal ions on enzyme activity.
Glucose 6-phosphate dehydrogenase (d-glucose 6-phosphate: NADP(+) oxidoreductase, EC 1.1.1.49; G6PD) is a key enzyme that is localized in all mammal tissues, especially in cytoplasmic sections and that catalyzes the first step of pentose phosphate metabolic pathway. In this study, G6PD enzyme was purified 1444-fold with a yield of 77% from rainbow trout liver using 2',5'-ADP-sepharose-4B affinity chromatography. Moreover, a purity check of the enzyme was performed with sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Some characteristic features like optimal pH, stable pH, optimal temperature and optimal ionic strength were determined for the purified enzyme. In addition to this, in vitro effects of ions like silver nitrate (Ag(+)), thallium sulphate (TI(+)), cobalt (II) nitrate (Co(2+)) and arsenic (V) oxide (As(5+)) on enzyme activity were researched. Half-maximal inhibitory concentration (IC50) values of Ag(+), Co(2+) and As(5+) metal ions, which showed an inhibitory effect, were found to be 0.0044, 0.084 and 4.058 mM, respectively; and their inhibition constants (K i) were found to be 0.0052 ± 0.00042, 0.087 ± 0.015700 and 4.833 ± 1.753207 mM, respectively. Tl(+) not exhibited inhibitory effect on the enzyme activity.
Agri Ibrahim Cecen University, Health Services Vocational School, Agri, Turkey.
Veysel
V
Comakli
Department of Chemistry, Faculty of Science, Ataturk University, Erzurum, Turkey.
Ebru
E
Akkemik
Department of Chemistry, Faculty of Science, Ataturk University, Erzurum, Turkey ciftcim@atauni.edu.tr.
Mehmet
M
Ciftci
Department of Chemistry, Faculty of Science, Ataturk University, Erzurum, Turkey.
Omer Irfan
OI
Kufrevioglu
D000818:Animals; D001151:Arsenic / Q000378:metabolism; D002846:Chromatography, Affinity; D003035:Cobalt / Q000378:metabolism; D004591:Electrophoresis, Polyacrylamide Gel; D004912:Erythrocytes / Q000187:drug effects / Q000378:metabolism; D029941:Fish Proteins / Q000737:chemistry / Q000302:isolation & purification; D005954:Glucosephosphate Dehydrogenase / Q000737:chemistry* / Q000302:isolation & purification*; D006863:Hydrogen-Ion Concentration; D007477:Ions; D008099:Liver / Q000201:enzymology*; D017686:Oncorhynchus mykiss / Q000378:metabolism*; D009994:Osmolar Concentration; D012835:Silver Nitrate / Q000378:metabolism; D013696:Temperature; D013793:Thallium / Q000378:metabolism
D029941:Fish Proteins; D007477:Ions; D003035:Cobalt; D012835:Silver Nitrate; D013793:Thallium; D005954:Glucosephosphate Dehydrogenase; D001151:Arsenic
Glucose 6-phosphate dehydrogenase; inhibition; liver; metal ion; rainbow trout