Experimental study of partitioning of 𝜶-Amylase in aqueous two phase systems based on Poly (ethylene) glycol 0222/ Magnesium sulfate

The aqueous two phase systems (ATPS) are introduced for separation of cells, amino acids, proteins and enzymes. In this study, the ATPS based on poly (ethylene glycol) 2000 and magnesium sulfate are applied for separation of α-amylase. The bimodal curve is firstly obtained by using cloud point method at 23 ℃ and is correlated by the Merchuk equation. Also, partitioning of α-amylase in two phase region at various working points is investigated and tie lines, partition coefficients and recovery are obtained. The results show that the bimodal curve is fitted with the Merchuk equation well. After partitioning of α-amylase enzyme in the ATPS, α-amylase tends to the top phase and the average of the partition coefficients the extraction recoveries are 1.15 and 71%, respectively. Also, the partition coefficient of α-amylase is increased with increasing concentration of polymer (at constant concentration of salt) and concentration of salt (at constant concentration of polymer). It is concluded that the aqueous two phase system based on poly (ethylene glycol) 2000 and magnesium sulfate can be used for separation of α-amylaseb as a suitable method.