CHIRAL STATIONARY PHASES BASED ON INTACT AND FRAGMENTED CELLOBIOHYDROLASE-I IMMOBILIZED ON SILICA

Cellobiohydrolase I (CBH I) was enzymatically degraded into two fragments which were immobilized on silica. Each fragment was shown to contain at least one enantioselective site for propranolol. The dominating enantioselective site of propranolol and other solutes is located on the main part of the enzyme -the core (C). CBH I was immobilized via its carboxylic groups onto aminopropyl silica and via its amino groups to aldehyde silica. The CBH I-aminopropyl silica separates enantiomers with higher enantioselectivity than does the CBH I-aldehyde silica, whereas the latter phase promotes high retention. Slow adsorption-desorption kinetics dominates the band broadening process of solutes on CBH I-silicas. Addition of dimethyloctylamine to the mobile phase resulted in a significant improvement of the peak efficiency of beta-blocking agents.