Abstract
The increasing demand for safe and effective methods for the separation of biological materials combining the advantageous properties of polyethylene glycol (PEG) (1500 g mol−1 or 4000 g mol−1) and ionic liquid has led to attractive and promising applications of the new aqueous biphasic systems. In this work, the liquid–liquid extraction process with aqueous two-phase systems (ATPS) was studied for α-amylase partitioning. The phase diagrams composed of PEG (1500 or 4000) g mol−1 + 1-ethyl-3-methylimidazolium chloride + water, were determined experimentally by the turbidimetric method at different temperatures T = (293.15, 303.15, 313.15 and 323.15) K. The effect of temperature and polymer molar mass on the phase diagrams, as well as the phase densities, refractive indexes and electrical conductivity were evaluated. Subsequently, the α-amylase partitioning was performed, where the partition coefficient was determined, besides the activity and selectivity. It was found that a temperature decrease and a polymer molar mass increase favored the formation of two phases. It was also found that α-amylase showed a preference for the bottom phase composed of PEG and the systems composed of PEG 1500 presented better results.
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Abbreviations
- ATPS:
-
Aqueous two-phase systems
- PEG:
-
Polyethylene glycol
- [C2min]Cl:
-
1-Ethyl-3-methylimidazolium chloride
- TLL :
-
Tie-line lengths
- STL :
-
Tie-line slope
- U:
-
One enzyme activity unit releases 1 mmol of reducing sugar per mL of extract per minute, U mL−1
- ART:
-
Total reducing sugars produced in the hydrolysis step, mg mL−1
- VT :
-
Total volume used in the hydrolysis, mL
- Vc :
-
Volume of sample used in the hydrolysis, mL
- Rt :
-
Reaction time, min
- BSA :
-
Bovine serum albumin
- K p :
-
Protein partition coefficient
- Ct :
-
Equilibrium concentration of the total protein in the top, mg mL−1
- Cb :
-
Equilibrium concentration of the total protein in the bottom, mg mL−1
- Ke :
-
Partition coefficient for enzyme activity
- At :
-
Enzyme activity in the top, mg mL−1
- Ab :
-
Enzyme activity in the bottom, mg mL−1
- \({\Delta }_{tr}G\) :
-
Gibbs free energy variation, kJ mol−1
- T :
-
Absolute temperature, K
- R :
-
Ideal gas constant, kJ mol−1 K−1
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Gandolfi, O.R.R., Gonçalves, G.R.F., Batista, I.C. et al. Aqueous two-phase system (polyethylene glycol + ionic liquid) for extraction of α-amylase: phase diagrams, systems characterization and partition study. Braz. J. Chem. Eng. 37, 595–606 (2020). https://doi.org/10.1007/s43153-020-00043-9
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DOI: https://doi.org/10.1007/s43153-020-00043-9