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

The HPLC separation of lignin peroxidase isoenzymes using Convective Intera ction Media disks containing quaternary amine and diethylaminoethyl ion-exc hange active groups is proposed. In contrast to standard HPLC procedures th e separation can be performed within a few minutes without considerably aff ecting the separation resolution. The method is reproducible and gives a li near response of integrated peak area to protein concentration for all meas ured isoenzymes. The separation resolution is retained unchanged by applyin g crude culture filtrate instead of a sample previously frozen and dialyzed . The optimized method might therefore be used for on-line monitoring of li gnin peroxidase isoenzyme composition during fermentation. On the other han d, the proposed method is comparable in time to the original method of lign in peroxidase activity measurement (proposed by Tien and Kirk), providing a dditionally the isoenzyme composition. (C) 1999 Academic Press.