A method of fast separation of lignin peroxidases using convective interaction media disks

Anal Biochem. 1999 Jul 15;272(1):43-7. doi: 10.1006/abio.1999.4146.

Abstract

The HPLC separation of lignin peroxidase isoenzymes using Convective Interaction Media disks containing quaternary amine and diethylaminoethyl ion-exchange active groups is proposed. In contrast to standard HPLC procedures the separation can be performed within a few minutes without considerably affecting the separation resolution. The method is reproducible and gives a linear response of integrated peak area to protein concentration for all measured isoenzymes. The separation resolution is retained unchanged by applying crude culture filtrate instead of a sample previously frozen and dialyzed. The optimized method might therefore be used for on-line monitoring of lignin peroxidase isoenzyme composition during fermentation. On the other hand, the proposed method is comparable in time to the original method of lignin peroxidase activity measurement (proposed by Tien and Kirk), providing additionally the isoenzyme composition.

Publication types

  • Comparative Study

MeSH terms

  • Chromatography, High Pressure Liquid / instrumentation
  • Chromatography, High Pressure Liquid / methods*
  • Chromatography, High Pressure Liquid / statistics & numerical data
  • Evaluation Studies as Topic
  • Isoelectric Focusing
  • Isoelectric Point
  • Isoenzymes / chemistry
  • Isoenzymes / isolation & purification*
  • Peroxidases / chemistry
  • Peroxidases / isolation & purification*
  • Phanerochaete / enzymology
  • Reproducibility of Results

Substances

  • Isoenzymes
  • Peroxidases
  • lignin peroxidase