Mikrobiyal Dipeptidil Peptidaz IV’ün Kinetik Özelliklerinin Belirlenmesi

Year 2018, , 28 - 37, 30.04.2018

Sevda Arısoy Özlem Üstün Aytekin

https://doi.org/10.13002/jafag4383

Abstract

Dipeptidil peptidaz IV (EC 3.4.14.5; DPPIV) enzimi tüm doku, organ ve sistemlerin düzenlenmesinde rol alan, nörolojik, immünolojik ve gastroenterolojik açıdan araştırılan önemli bir enzimdir. DPPIV enzimi pek çok memeli, bitki, böcek ve mikrobiyal hücrelerden regüle edici enzim olarak elde edilmektedir. DPPIV enziminin sindirim enzimi eksikliği, otizm, çölyak ve Asperger sendromu gibi hastalıklarda gıda takviyesi olarak kullanımıyla son yıllarda ticari olarak üretilmesi artmıştır. DPPIV enzim kaynağı olarak hücre-dışı enzim ekspresyonu yapan fungal kaynaklar tercih edilmesine rağmen bakterilerin enzim üretim verimliliği funguslara göre daha fazla olduğundan son yıllardaki ticari enzim üretimlerinde bakteriler tercih edilmektedir. Bu çalışmada, koleksiyon kültüründen satın alınmış bir laktik asit bakterisi, Lactococcus lactis spp. lactis (L. lactis), enzim üreticisi mikroorganizma olarak seçilmiştir. L. lactis, kazein proteinini hidrolizlemek için endo- ve ekzo-peptidazlar eksprese eder. DPPIV enzimi de bunlardan biridir. Hücre-içi DPPIV enzimi, üretiminden sonra saflaştırılmış ve ardından enzim karakterizasyonunu yapmak için kinetik özellikleri incelenmiştir. Sonuçlar, Gly-Pro-pNA substratı için optimum sıcaklığın 40 °C ve optimum pH’ nın 8,0 olduğunu göstermektedir. Saflaştırılan enzimin Gly-Pro-pNA substratına ilgisi olduğu bulunmuştur. Enzim, Hg+2, Zn+2, Ca+2, Fe+2 ve Mg+2 iyonları tarafından sırası ile inhibe edilmiştir. Enzim, Diprotin A tarafından güçlü bir şekilde inhibe edilmiştir. Jelatin inhibisyonu fenilmetilsulfonil florid (PMSF) inhibisyonu ile yakın bulunmuş ve etilendiamin tetra asetik asitin (EDTA) inhibisyona etkisi gözlenmemiştir.

Keywords

Dipeptidil Peptidaz IV, enzim inhibisyonu, enzim kinetiği, Lactococcus lactis spp. lactis

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