USES OF BETA-GALACTOSIDASE TAG IN ONLINE MONITORING PRODUCTION OF FUSION PROTEINS AND GENE-EXPRESSION IN ESCHERICHIA-COLI

A simple method for monitoring and quantifying automatically the production by fermentation of beta-galactosidase fusion proteins, making use of the remaining activity of the beta-galactosidase part, is considered. A hybrid protein carrying the major antigenic domain of foot-and-mouth disease virus C1 joined at the N-terminus of beta-galactosidase has been expressed in Escherichia coli. The yield of the chimeric protein has been monitored by flow injection analysis (FIA) during batch fermentations at laboratory scale, and a high correlation between values of product concentration from FIA and from immunological quantizations has been obtained. Because of the possibility of employing FIA in large-scale experiments, and the high sampling frequency, versatility, and reproducibility offered by this method, we propose FIA as a general, simple, quick, flexible, and reliable instrument for both monitoring the yield of recombinant proteins produced industrially, and performing basic research at laboratory scale.