Comakli Veysel, Akkemik Ebru, Ciftci Mehmet, Kufrevioglu Omer Irfan
Agri Ibrahim Cecen University, Health Services Vocational School, Agri, Turkey.
Department of Chemistry, Faculty of Science, Ataturk University, Erzurum, Turkey.
Toxicol Ind Health. 2015 May;31(5):403-11. doi: 10.1177/0748233713475514. Epub 2013 Jan 30.
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.
葡萄糖-6-磷酸脱氢酶（D-葡萄糖-6-磷酸：NADP(+)氧化还原酶，EC 1.1.1.49；G6PD）是一种关键酶，存在于所有哺乳动物组织中，尤其是细胞质部分，它催化磷酸戊糖代谢途径的第一步。在本研究中，使用2',5'-ADP-琼脂糖-4B亲和色谱法从虹鳟鱼肝中纯化出G6PD酶，纯化倍数为1444倍，产率为77%。此外，用十二烷基硫酸钠-聚丙烯酰胺凝胶电泳对该酶进行了纯度检查。测定了纯化酶的一些特性，如最适pH、稳定pH、最适温度和最适离子强度。除此之外，还研究了硝酸银（Ag(+)）、硫酸铊（TI(+)）、硝酸钴（II）（Co(2+)）和五氧化二砷（As(5+)）等离子对酶活性的体外影响。发现具有抑制作用的Ag(+)、Co(2+)和As(5+)金属离子的半最大抑制浓度（IC50）值分别为0.0044、0.084和4.058 mM；它们的抑制常数（Ki）分别为0.0052±0.00042、0.087±0.015700和4.833±1.753207 mM。Tl(+)对酶活性未表现出抑制作用。
