Alkalen Fosfataz (Alp) Enziminin Özütlenmesi, İnhibisyonu Ve Kinetik Modellenmesi

Öz

Bu çalışma, sıçan karaciğerinden özütlenen alkalen fosfataz (ALP) enziminin kısmen özütlenmesini, inhibitörsüz ve inhibitör varlığında kinetik özelliklerinin araştırılmasını kapsamaktadır. Deneysel sonuçlar yardımıyla Michaelis Menten ve Lineweaver Burk grafikleri çizildi. Bu grafiklerden Km ve Vmax değerleri hesaplandı. Değerler: İnhibitörsüz Km= 0,047 Vmax=24,87; Üre inhibitörlü Km= 0,054 Vmax=20,83 ; Kreatinin inhibitörlü Km= 0,037 Vmax=20,83 olarak hesaplanmıştır. Bu değerlere göre enzimin substrata ilgisi ve maksimum hız tespit edildi. İnhibitör maddeyle deneyler tekrarlandı ve inhibisyon tipi belirlendi. Kandaki yüksek üre ve kreatinin seviyesinin alkalen fosfataz enzimini nasıl etkilediği bulundu. Kandaki ALP (Alkalen Fosfataz) üzerinde ürenin
inhibitör olarak kullanıldığı çalışmada reaksiyonun yarışmasız (nonkompetetif) bir reaksiyon, kreatininin inhibitör olarak kullanıldığı çalışmada ise reaksiyonun yarışmacı olmayan (unkompetetif) bir reaksiyonla gerçekleştiği tespit edilmiştir.

Anahtar Kelimeler

• Alkalen fosfataz, Michaelis Menten, yarışmasız inhibisyon, yarışmacı olmayan inhibisyon.

ISOLATION, INHIBITION AND KINETIC MODELLING OF ALKALINE PHOSPHATASE (ALP) ENZYME

Öz

This study scopes the purification of alkaline phosphatase enzyme extracted from the liver and the investigation
of kinetic properties for the reactions with and without inhibitor. Michaelis Menten and Lineweaver Burk graphs
were drawn by means of experimental results. Km and Vmax values were calculated from these graphics.
According to these values, substrate affinity (Km) and maximum velocity (Vmax) values were determined. Values
without inhibitor; Km= 0,047 Vmax=24,87; with urea inhibitor Km= 0,054 Vmax=20,83 ; with creatinin inhibitor
Km= 0,037 Vmax=20,83. The experiments were repeated with inhibitory substance and inhibition type was
established. How high blood urea and creatinine levels affected alkaline phosphatase activity were determined.
In this study, urea was found to cause noncompetetive inhibition and creatinine to cause uncompetetive
inhibition on liver ALP. 

Anahtar Kelimeler

• Alkaline phosphatase,Michaelis Menten,Lineweaver Burk,noncompetetive inhibition,uncompetetive inhibition

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