Separation of manganese peroxidase isoenzymes on strong anion-exchange monolithic column using pH-salt gradient.
Different chromatographic methods including chromatofocusing are used for separation of manganese peroxidase (MnP) isoforms and their isolation from the fungal growth medium. We tested strong anion exchange methacrylate based monolithic columns as a stationary phase for fast separation of MnP's. Sodium acetate buffers of two different pH values (6 and 4) were used for formation of reproducible pH gradient. The entire cycle, involving analysis and column regeneration, was completed in 3 min. Use of pH gradient showed better MnP isoform separation comparing to the salt gradient, while application of combined pH-salt gradient, resulted in further improvement.
Podgornik
Helena
H
University of Ljubljana, Faculty of Chemistry and Chemical Technology, Askerceva 5, Ljubljana 1000, Slovenia.
Podgornik
Ales
A
D000837:Anion Exchange Resins; D002851:Chromatography, High Pressure Liquid / Q000295:instrumentation* / Q000379:methods; D006863:Hydrogen-Ion Concentration; D007525:Isoelectric Focusing; D007527:Isoenzymes / Q000302:isolation & purification*; D010544:Peroxidases / Q000302:isolation & purification*; D020075:Phanerochaete / Q000201:enzymology; D012492:Salts / Q000737:chemistry
D000837:Anion Exchange Resins; D007527:Isoenzymes; D012492:Salts; D010544:Peroxidases; C051129:manganese peroxidase