Factors Affecting the Partitioning of Peroxidase from Aqueous Two-Phase Extraction
Keywords:
Peroxidase, Aqueous Two-Phase Extraction, Partitioning, CabbageAbstract
Peroxidase is an enzyme that catalyzes the oxidation of various substances by utilizing hydrogen peroxide as an electron acceptor. It plays a significant role in multiple aspects, including laboratory analysis and immunoassays. Peroxidase is found abundantly in plant. Therefore, the purpose of this research is to study the factors affecting the partitioning of peroxidase using aqueous two-phase extraction (ATPE) consisting of polyethylene glycol (PEG) and inorganic salts. Cabbage was chosen as a source of peroxidase enzyme. The results showed that changes in the type of salt used to build the system (ammonium sulfate ((NH4)2SO4), sodium sulfate (Na2SO4), dipotassium hydrogen phosphate (K2HPO4) and disodium hydrogen phosphate (Na2HPO4)), molecular weight of polyethylene glycol (PEG 1500 and 6000), polyethylene glycol concentration (19–31 %w/v), salt concentration (10–22 %w/v), system pH (4–8), and the effect of sodium chloride ( 0–5%w/v) affected the partition coefficient of the enzyme (KE), the partition coefficient of the protein (KP), volume fraction of phase (Vr), the percent yield (%Yield) and the purification factor (PF). The optimum condition for peroxidase extraction obtained in this study was using ammonium sulfate salt at a concentration of 16%w/v, polyethylene glycol molecular weight 1500 at a concentration of 25%w/v, the pH of the system was 6 and no sodium chloride was added. The partition coefficient of the enzyme was 15.75 ± 4.07, the percent yield was 97.06 ± 0.71, and the purification factor was 4.86 ± 0.70 times.
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