A technique based on liquid chromatography has been developed to facilitate studies of enantioselectivity in penicillin G acylase (PGA)-catalyzed hydrolysis of some 2-aryloxyalkanoic acid methyl esters and isosteric analogues. PGA was covalently immobilized on an aminopropyl monolithic silica support to create an immobilized HPLC-enzyme reactor. Two sets of experimental data were drawn to calculate the enantioselectivity (E) of the kinetically controlled enantiomer-differentiating reaction, the degree of substrate conversion and the enantiomeric excess of the product. The developed enzymatic reactor was coupled through a switching valve to an achiral analytical column for separation and quantitation of the hydrolysis products. The enantiomeric excess was determined off-line on a PGA-chiral stationary phase. In this way, highly precise E values were determined. A computational study related to the hydrolysis of the considered racemic esters was also carried out in order to unambiguously clarify both the substrate specificity and the enantioselectivity displayed by PGA.
Enantioselective Hydrolysis of Some 2-Aryloxyalkanoic Acid Methyl Esters and Isosteric Analogs Using a Penicillin G Acylase Based HPLC Monolithic Silica Column / Massolini, G.; Calleri, E.; Lavecchia, Antonio; Loiodice, F.; Lubda, D.; Temporini, C.; Fracchiolla, G.; Tortorella, P.; Novellino, Ettore; Caccialanza, G.. - In: ANALYTICAL CHEMISTRY. - ISSN 0003-2700. - STAMPA. - 75:(2003), pp. 535-542.
Enantioselective Hydrolysis of Some 2-Aryloxyalkanoic Acid Methyl Esters and Isosteric Analogs Using a Penicillin G Acylase Based HPLC Monolithic Silica Column
LAVECCHIA, ANTONIO;NOVELLINO, ETTORE;
2003
Abstract
A technique based on liquid chromatography has been developed to facilitate studies of enantioselectivity in penicillin G acylase (PGA)-catalyzed hydrolysis of some 2-aryloxyalkanoic acid methyl esters and isosteric analogues. PGA was covalently immobilized on an aminopropyl monolithic silica support to create an immobilized HPLC-enzyme reactor. Two sets of experimental data were drawn to calculate the enantioselectivity (E) of the kinetically controlled enantiomer-differentiating reaction, the degree of substrate conversion and the enantiomeric excess of the product. The developed enzymatic reactor was coupled through a switching valve to an achiral analytical column for separation and quantitation of the hydrolysis products. The enantiomeric excess was determined off-line on a PGA-chiral stationary phase. In this way, highly precise E values were determined. A computational study related to the hydrolysis of the considered racemic esters was also carried out in order to unambiguously clarify both the substrate specificity and the enantioselectivity displayed by PGA.I documenti in IRIS sono protetti da copyright e tutti i diritti sono riservati, salvo diversa indicazione.
