Separation of invertase enzyme from tomato using aqueous two-phase system: PEG1500/sodium citrate and PEG4000/sodium citrate
DOI:
https://doi.org/10.31699/IJCPE.2026.3.4Keywords:
Aqueous two-phase system; Enzymes; plant source; extractionAbstract
An aqueous two-phase system (ATPS) is a highly useful, effective, simple, and economically friendly technique for the biomolecules recovery. ATPS is created by mixing two polymers or a polymer and salt above their critical concentration in water. Invertase is an enzyme that hydrolyzes sucrose into glucose and fructose, making it highly crucial for the food industry. Because the ATPS can be applied in mild conditions, in a water-rich phase, and is biocompatible with bioproducts, this method was used to extract the invertase enzyme from tomato pulp. Specific activity and protein concentration of the enzyme were found to be 10.87 U/mg and 9.44 mg/mL, respectively. In this research, two ATPS systems were used: polyethylene glycol 1500/sodium citrate (PEG1500/SC) and polyethylene glycol 4000/sodium citrate (PEG4000/SC). Five parameters were studied to obtain the maximum amount of invertase from tomato: temperature, pH, concentrations of PEG and salt, and supporting neutral salt (NaCl and MgCl2). The maximum percent recovery (%Rec) of invertase and its partition coefficient (KE) were 84% and 5.25 respectively, at a temperature of 10 oC, pH 10, and concentrations of PEG1500 and SC of 1.8 g/10 ml each for system-1 (PEG1500/SC) without adding a supporting neutral salt, whereas for system-2 (PEG4000/SC), the optimum operating conditions were at a temperature of
10 oC, pH of 10, and concentrations of PEG4000 and SC of 1.65 g/10 ml and 1.5 g/10 ml respectively, where the maximum %Rec and KE were 89.8% and 8.8, respectively, without adding supporting neutral salt. Because system-2 provides better results than system-1, it is considered to improve the enzyme recovery by adding NaCl or MgCl2 as supporting neutral salts, where the %Rec increased to 95.8% and 96.7% for the NaCl and MgCl2 additions, respectively.
Received on 24/05/2026
Received in Revised Form on 26/06/2026
Accepted on 26/06/2026
Published on 30/09/2026
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