Abstract
Preparative isoelectric focusing of grapevine peroxidases on glycerol-stabilized pH gradients in the RotoforTM cell has been used to purify peroxidase isoenzymes from the spent medium of suspension cultured cells derived from mesocarp tissues of grape (Vitis vinifera cv. Monastrell) berries. The results showed that, under steady-state conditions, basic peroxidase isoenzymes focus well while acidic peroxidase isoenzymes show anomalous behaviour during isoelectric focusing, leading to diffuse and/or multiband isoelectric focusing patterns. These results suggest that, although preparative isoelectric focusing in glycerol-stabilized pH gradients using the Rotofor cell may be a powerful tool for isoenzyme purification, the advantages of this technique may be severely hampered by the anomalous behaviour shown by acidic peroxidase isoenzymes during focusing.
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