Expression and purification of recombinant phenylalanine 2,3-aminomutase from Pantoea agglomerans

In the present study, the gene of phenylalanine 2,3-aminomutase from Pantoea agglomerans (PaPAM) was cloned into pET-19b vector and used for its expression in competent Escherichia coli cells. The recombinant plasmid, PaPAM-pET-19b, was transformed into competent E. coli strain BL21(DE3) pLysS cells. Overnight culture of the transformed bacteria was induced by the addition of isopropylthio-beta-D-galactoside (IPTG) to the final concentrations of 0.1, 0.5 and 1 mM. Also, the effects of different temperatures (18, 25 and 30 degrees C) and the incubation time of PaPAM were examined. The fermentation process was scaled up to 10 L fermentor. Affinity purification conditions were analyzed by SDS-PAGE. The T-m and the activity of the purified enzyme was also investigated.