Koyun Karaciğerinden Alkol Dehidrogenaz: Saflaştırma, Karakterizasyon, ve Bazı Antibiyotiklerin Etkileri

Year 2017, Volume: 7 Issue: 3, 151 - 160, 30.09.2017

Yeliz Demir Bülent Şengül Bülent Ergun Şükrü Beydemir

Abstract

Alkol dehidrogenaz (ADH), her bir alt biriminin bir Zn2+ metal içeren katalitik alana ve bir kofaktör bağlama alanına sahip olduğu dimerik bir enzimdir. Bu enzim, alkolü, aldehide dönüştürür. Bu makale koyun karaciğerinden alkol dehidrogenazın saflaştırılması, karakterizasyonu enzimin aktivitesi üzerine bazı antibiyotiklerin in vitro ekilerine odaklanmaktadır. ADH, DEAE-Sephadex A-50 iyon değişim kromatografsi ve Sephadex G-100 jel fltrasyon kromatografsi vasıtasıyla koyun karaciğerinden 0.5 U mg-1 protein spesifk aktivite ve yaklaşık olarak 52.03-kat saflaştırıldı. Enzimin alt birim ve doğal hallerinin molekül kütleleri jel fltrasyon kromatografsi ve SDS-PAGE ile sırasıyla 38.16 kDa ve 80.49 kDa olarak belirlendi. ADH’ın optimum iyonik şiddeti, sıcaklığı ve pH’sı sırasıyla 400 mM, 40°C ve 10.5 idi. Antibiyotiklerin inhibitor etkileri çeşitli konsantrasyonlarda denendi. Kanamisin sülfat, amikasin sülfat, gentamisin, linkomisin ve klindamisin için IC50 değerleri sırasıyla, 43.31, 36.47, 20.38, 18.73 ve 1.31 mM olarak bulundu.

Keywords

Alkol dehidrogenaz, antibiotikler, inhibisyon, karakterizasyon, saflaştırma

Alcohol Dehydrogenase from Sheep Liver: Purification, Characterization and Impacts of Some Antibiotics

Abstract

Alcohol dehydrogenase (ADH) is a dimeric enzyme in which each subunit of the enzyme has a Zn2+ metal-containing catalytic domain and a cofactor-binding domain. This enzyme converts the alcohol to aldehyde. The present article focuses on the purification, characterization and in vitro effects of some antibiotics on alcohol dehydrogenase from sheep liver. ADH was purified with specific activity of 0.5 U/mg proteins and approximately 52.03-fold from sheep liver by DEAE-Sephadex A-50 ion exchange chromatography and gel filtration on Sephadex G-100. The subunit and the natural molecular weights of the enzyme were determined by gel filtration and SDSPAGE 38.16 kDa and 80.49 kDa, respectively. The optimum ionic strenght, temperature and pH of ADH were found 400 mM, 40 °C and 10.5, respectively. The inhibitory effects of the antibiotics were tested at various concentrations. IC50 values for kanamycin sulfate, amikacin sulfate, gentamicin, lincomycin, and clindamycin were found to be 43.31, 36.47, 20.38, 18.73 and 1.31 mM, respectively

Keywords

Alcohol dehydrogenase, antibiotics, characterization, inhibition, purifcation

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