Orhan ERDOĞAN, Abdulkadir ÇİLTAŞ, Olcay HİSAR, Mehmet ÇİFTÇİ

Inhibition effects of some antibiotics on the activity of glucose 6- phosphate dhydrogenase enzyme from rainbow trout (Oncorhynchus mykiss Valbaum, 1792) erythrocytes

Bazı antibiyotiklerin, gökkuşağı alabalığı (Oncorhynchus mykiss Waibaum, 1792) eritrositlerindeki glikoz 6-fosfat dehidrogenaz enzimi üzerine inhibisyon etkileri araştırılmıştır. Bu amaçla başlangıçta, eritrositlerdeki glukoz 6-fosfat dehidrogenaz enzimi, amonyum sülfat çöktürmesi ve 2',5' Sefaroz 4B afinite jeli kullanılarak, %70,40 oranında 1271.19 kat saflaştırılmıştır. Saflaştırma esnasında sıcaklık +4 °C' de sabit tutulmuş, enzim aktivitesi spektofotometre ile 340 nm'de Beutler metoduna göre ölçülmüştür. Bütün kinetic çalışmalarında bu metot kullanılmıştır. Antibiyotiklerden; tiamfenikol, amikasin, gentamisin ve netilmisin kullanılmıştır. Bu antibiyotiklerin hepsi enzim üzerinde inhibitor etki göstermiştir. Glukoz 6-fosfat dehidrogenaz enzimi için K: değeri Lineweaver-Burk grafiklerinden tespit edilmiştir. Tiamfenikol ve amikasin yarışmalı bir inhibisyon gösterirken, gentamisin ve netilmisin yarışmasız bir inhibisyon göstermiştir. Buna ilaveten, antibiyotiklerin I50 değerleri, % vs [1] aktivitesinden tayin edilmiştir.

Gökkuşağı alabalığı (Oncorhynchus mykiss Walbaum, 1792) eritrositlerindeki glikoz 6-fosfat dehitrogenaz enzimi aktivitesi üzerine bazı antibiyotiklerin inhibisyon etkileri

Inhibitory effects of some antibiotics on glucose 6-phosphate dehydrogenase from the erythrocytes of rainbow trout (Oncorhynchus mykiss Waibaum, 1792) were investigated. For this purpose, initially erythrocyte glucose 6-phosphate dehydrogenase was purified 1271.19-fold in a yield of 70.40% by using ammonium sulfate precipitation and 2',5'-ADP Sepharose 4B affinity gel. A constant temperature (+4 °C) was maintained during the purification process. Enzyme activity was determined with the Beutler method by using a spectrophotometer at 340 nm. This method was utilized for all kinetic studies. Thiamphenicol, amikacin, gentamicin, and netilmicin were used as antibiotics. All the antibiotics exhibited inhibitory effects on the enzyme. K, constants for glucose 6-phosphate dehydrogenase were found by means of Lineweaver-Burk graphs. While thiamphenicol and amikacin showed competitive inhibition, gentamicin and netilmicin displayed noncompetitive inhibition, in addition, lso values of the antibiotics were determined by plotting activity % vs. [1].

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