Purification of peroxidase from Amsonia orientalis by three-phase partitioning and its biochemical characterization

ABSTRACT

The present work describes the purification and characterization of peroxidase from the medicinal plant, Amsonia orientalis, for the first time. The activity recovery for peroxidase was 162% with 12.5-fold purification. Optimal purification parameters were 20% (w/v) (NH₄)₂SO₄ saturation at pH 6.0 and 25°C with 1.0:1.0 (v/v) ratio of crude extract to t-butanol ratio for 30 min. The molecular mass of the enzyme was found to be ca. 59 kDa. Peroxidase showed K_m values of 1.88 and 2.0 mM for pyrogallol and hydrogen peroxide, respectively. FeSO₄, CuSO₄, HgCl₂, MnSO₄ and MgSO₄ did not inhibit the enzyme activity.

KEYWORDS:

• Activity recovery
• molecular weight
• peroxidase
• purification
• three-phase partitioning

Funding

We would like to thank The Scientific and Technological Research Council of Turkey (Grant number: 113Z609) and Kocaeli University (Project number: 2017/057) for funding.

Additional information

Funding

We would like to thank The Scientific and Technological Research Council of Turkey (Grant number: 113Z609) and Kocaeli University (Project number: 2017/057) for funding.