Three-Phase Partitioning of α-Galactosidase from Aspergillus lentulus: Optimization of System and Characterization of Enzyme

Year 2020, Volume: 48 Issue: 1, 83 - 98, 17.04.2020

Derya Çamurlu Hasan Bayraktar , Sennur Çalışkan Ataç Uzel , Seçil Önal

https://doi.org/10.15671/hjbc.511731

Cited By: 1

Abstract

Üçlü-faz ayırma
(TPP) tekniği α-galaktozidazın Aspergillus lentulus’dan tek adımda kısmi
saflaştırılması için ilk kez başarıyla kullanıldı. Yüksek aktivite ve
saflaştırma katı elde etmek için enzimin ekstraksiyon etkinliğine amonyum
sülfat konsantrasyonu, ekstrakt t-butanol oranı ve pH etkisi araştırıldı.
Sistemin optimum saflaştırma parametreleri %55(w/v) amonyum sülfat
konsantrasyonu, 1:1 (v/v) ham ekstrakt t-butanol oranı ve pH 5.5 olarak
belirlendi. Bu optimize TPP sistemi α-galaktosidaz için 5.3 saflaştırma
katı ile %178 aktivite verimi oluşturdu. pH 6.5 ve 50^oC’de maksimum aktivite gözlendi. α-Galaktozidaz 25-60°C sıcaklık aralığında ve pH 2.6-5.5 aralığında
oldukça iyi bir kararlılık gösterdi. K_M ve V_max değerleri
sırasıyla 0.365 mM ve 0.093 U olarak belirlendi. Metal iyonları ve şekerler
arasında Na₂CO₃ ve
galaktoz enzim aktivitesi üzerinde kuvvetli inhibitor etkisi gösterdi. TPP ile A. lentulus’dan farklı biyokimyasal
özelliklere sahip yeni bir α-galaktozidazın elde edilmesi onun çeşitli biyoteknolojik
uygulamaları açısından ilgi çekici olacaktır. 

Keywords

α-galactosidase, bioseparation, three-phase partitioning (TPP), enzyme purification

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