The Use of Monolithic Columns for the Separation of Potato Protease Inhibitors Using the Fast Protein Liquid Chromatography System
Department of Special Plant Production, Faculty of Agriculture, University of South Bohemia, České Budějovice, Czech Republic
Keywords: fast protein liquid chromatography, potatoes, protease inhibitors, SDS-PAGE
Juices from tubers of two potato varieties, Adéla and Eurostarch, have been selected as the source material for the separation of protease inhibitors. Their purification was achieved first using the gravity flow ion-exchange and Con-A affinity columns followed by the chromatographic separation applying monolithic ion exchange columns attached to a Fast Protein Liquid Chromatography system. Protease inhibitors were identified in separated fractions by SDS-PAGE. Potato inhibitor II and potato inhibitor I were major components in acidic fractions. The alkaline protease inhibitors were identified as isoforms of cysteine protease inhibitors, aspartate protease inhibitors, and as some proteins of the complex Kunitz-type protease inhibitors family.
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