Evaluation of a monolithic epoxy silica support for penicillin G acylase immobilization.
In this work, a new type of penicillin G acylase (PGA)-based monolithic silica support was developed and evaluated for the chiral separation in HPLC. The preparation procedure consisted of two steps: preparation of an epoxy derivatized monolithic silica column and chemical modification of the epoxide groups with the enzyme chiral selector. The epoxy Silica-Rod column for the immobilization of PGA was prepared with the in situ modification process by using epoxy-silanes and the identification of the species bound to the surface was achieved by solid-state nuclear magnetic resonance. The enzyme was covalently immobilized to the surface of the derivatized monolithic column. The enantioselectivity and the performance of the developed column are discussed and compared to the corresponding experimental data obtained with a PGA-based microparticulate (5 microm) silica column.
Calleri
E
E
Department of Pharmaceutical Chemistry, University of Pavia, Via Taramelli 12, 27100 Pavia, Italy. enrica.calleri@unipv.it
Massolini
G
G
Lubda
D
D
Temporini
C
C
Loiodice
F C
FC
Caccialanza
G
G
D002851:Chromatography, High Pressure Liquid; D004800:Enzymes, Immobilized / Q000737:chemistry*; D004852:Epoxy Compounds / Q000737:chemistry*; D007202:Indicators and Reagents; D009682:Magnetic Resonance Spectroscopy; D008863:Microspheres; D010316:Particle Size; D010399:Penicillin Amidase / Q000737:chemistry*; D012822:Silicon Dioxide / Q000737:chemistry*; D013056:Spectrophotometry, Ultraviolet; D013237:Stereoisomerism; D013499:Surface Properties
D004800:Enzymes, Immobilized; D004852:Epoxy Compounds; D007202:Indicators and Reagents; D012822:Silicon Dioxide; D010399:Penicillin Amidase