METHOD OF OBTAINING PEROXYDASE ENZYME FROM HORSE RADISH ROOTS Russian patent published in 2009 - IPC C12N9/02 

FIELD: chemistry, biochemistry.

SUBSTANCE: method includes crashing and homogenisation of sprouted horse radish roots, extraction of homogenisate of horse radish roots with 0.15±0.01 M solution of sodium chloride. Ballast proteins are separated and peroxidase is precipitated with ammonium sulphate salts of 45-48% saturation and 85-90% saturation respectively. Gel filtration of peroxidase solution is carried out on Sephadex G-100 with eluting with 0.5-0.2 M sodium chloride solution. Chromatographic purification is carried out on carboxymethylcellulose. Liophyl drying of target peroxidase is carried out. Dialysis against sodium-acetate buffer with pH 4.4-5.0 and dialysis against potassium-phosphate buffer with pH 8.0±0.1 is carried out. Concentration with additional purification on DEAE-cellulose with elution with the same buffer with further dialysis against deionised water is carried out. Method allows to obtain peroxidase with high specific activity and high output. Peroxidase output constitutes 2.52-3.50 g/kg of horse radish roots, specific activity is 640-700 A.U./mg protein.

EFFECT: obtaining peroxidase with high specific activity and high output.

6 cl, 2 dwg, 1 tbl, 2 ex