Abstract
The effect of poly(ethyleneglycol) (PEG) molecular weight, system pH, and sodium chloride concentration on the partitioning behavior of horseradish peroxidase fromArmomcia rusticana root extract was investigated in poly(ethyleneglycol)/sodium phosphate systems.
PEG molecular weight strongly affects the enzyme partition coefficient, whereas pH variation from 5.5 to 8.0 has little effect. The addition of sodium chloride (8% w/w) to a PEG 1540/phosphate system, pH 7.0, raises the peroxidase partition coefficient 13.5-fold without important changes in that of total horseradish root proteins. Moreover, these conditions allow direct homogenization of theA. rusticana roots with the selected aqueous two-phase system with the clear top phase containing over 90% of the enzyme and the purification factor being 4.8.
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Abbreviations
- ATPS:
-
aqueous two-phase systems
- PEG:
-
poly(ethyleneglycol)
- PO:
-
horseradish peroxidase
- TP:
-
total protein
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Miranda, M.V., Lahore, H.m.F. & Cascone, O. Horseradish peroxidase extraction and purification by aqueous two-phase partition. Appl Biochem Biotechnol 53, 147–154 (1995). https://doi.org/10.1007/BF02788604
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DOI: https://doi.org/10.1007/BF02788604