METHOD OF OBTAINING PEROXIDASE FROM HORSERADISH ROOT Russian patent published in 2013 - IPC C12N9/08 

FIELD: biotechnology.

SUBSTANCE: crushed biomass of horseradish root is soaked in 0.1 M sodium phosphate buffer solution pH 7.0, with preliminary blown nitrogen in presence of 5 mcM hemin solution and 5 mcM calcium chloride. The extract is separated by decantation followed by filtration and concentrating by ultrafiltration through ultrafilters with a pore size of less than 30 kilodalton. The enzyme extract is saturated with ammonium sulfate to 35% saturation and applied to a column with phenyl-sepharose after extensive washing of the buffer with sulfate, the active fractions are removed with a gradient of ammonium sulfate (35%-0%) and increasing the pH to 8.0. The enzyme is purified by gel filtration on Toyopearl HW55F, dialysed and dried lyophilisation.

EFFECT: use of hemin-containing buffers at a stage of extraction and gel filtration enables to obtain highly active enzyme with high acceleration of enzyme production process and improving its catalytic characteristics and stability.

7 cl, 1 tbl