Partition of horseradish peroxidase in aqueous two-phase systems containing polyvinylpirrolidone

M.V. MIRANDA; O. CASCONE

BIOSEPARATION

Kluwer Academic Publishers

Año: 1997 vol. 7 p. 25 - 30

0923-179X

Growing peroxidase utilisation in different industries encourages the search for high benefilcost ratio purification methods such as aqueous two-phase partition. In this way, the partitioning behaviour of peroxidase from Armoracía rusticana roots in polyvinylpinolidone/Reppal and polyvinylpinolidone/salt aqueous two-phase systems was investigated. Based on these results, a two-step purification process was developed. In the first system(polyvinylpyrolidone K3O/Reppal PES 200, pH 7.0), cell debris and some contaminating proteins were shifted to the bottom phase while peroxidase concentrated in the top phase. After discarding the bottom phase, the second step involved addition of magnesium sulphate thus forming a second aqueous two-phase system. At this step, the enzyme was extracted into the salt-rich bottom phase. The overall yield was 75% and the purification factor 7.3.