Arzu ÖZTÜRK KESEBİR, Deryanur KILIÇ, Ömer KÜFREVİOĞLU

Lipoksigenaz Enziminin Sığır Karaciğerinden Kısmi Saflaştırılması, Karakterizasyonu, Salisilik Asitin ve Bazı Flavonların Enzim Üzerine Etkileri

Lipoksigenaz (EC 1.13.11.34.; LOX ) enzimleri; iki ya da daha fazla doymamış bağ bulunduran yağ asitlerine oksijen katarak oksitleyen, yapısında hem grubu olmayan demir taşıyıcı dioksigenazlardır. Gelişmiş bitkilerde, memelilerde, ökoryatik alglerde, fungilerde, bazı bakterilerde de tanımlanmıştır. Hayvan dokularında, LOX 5, LOX 8, LOX 12 ve LOX 15 izoenzimlerinin bulunduğu doğrulanmıştır. Bu çalışmada, sığır karaciğerinden LOX enzimi % 30-50 doygunluk aralığında amonyum sülfat çöktürmesiyle kısmi saflaştırıldı. Karakterizasyon çalışmaları yapılarak, optimum iyonik şiddet değeri 0.5 mM, optimum pH değeri 4.0 ve optimum sıcaklık değeri 30 °C olarak belirlendi. LOX enzimi aktivitesi üzerinde salisilik asit, eupatorin, eupatilin ve gardenin A maddelerinin inhibisyon etkisini araştırmak üzere 5 farklı inhibitör konsantrasyonunda aktivite ölçümü yapılarak % Aktivite –[I] grafikleri çizildi, IC50 değerleri bulundu. Salisilik asit için 1.43 μM, eupatorin için 0.46 μM, eupatilin için 0.15 μM ve gardenin A için 5.31 μM olarak belirlendi.

Anahtar Kelimeler:

Lipoksigenaz, kısmi saflaştırma, karakterizasyon, inhibisyon

Partial Purification and Characterization of Lipoxygenase Enzyme From Bovine Liver, Effects of Salicylic Acid and Some Flavons on the Enzyme

Lipoxygenase (EC 1.13.11.34; LOX) is a non-hem iron-bearing dioxygenase which converts oxygen into fatty acids containing two or more unsaturated bonds and converts them into fatty acid hydroperoxides. Lipoxygenase activity is described in developed plants, in mammals, in echolalia, in fungi, in bacteria. LOX 5, LOX 8, LOX 12 and LOX 15, the four-headed isoenzymes of LOX enzymes are commonly found in animal tissues. In this study, LOX enzyme from bovine liver was partially purified by precipitation of ammonium sulphate at 30-50% saturation. Characterization studies were carried out and the optimum ionic strength value was determined as 0.5 mM, optimum pH value was pH 4.0 and optimum temperature value was 30 °C. The inhibitory effect of salicylic acid, eupatorin, eupatilin and gardenin A on LOX enzyme was investigated. Inhibitory effect of salicylic acid, eupatorin, eupatilin and gardenin A on LOX enzyme was investigated. IC50 values were calculated as 1.43 μM for salicylic acid, 0.46 μM for eupatorin, 0.15 μM for eupatilin and 5.31 μM for gardenin A.

Keywords:

Lipoxygenase, partial purification, characterization, inhibition,

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