Characteristics of the melibiose transporter and its primary structure in Enterobacter aerogenes

Abstract

Cells of Enterobacter aerogenes can grow on melibiose as a sole source of carbon. This suggests the presence of melibiose operon in this organism. We found that E. aerogenes cells possess both α-galactosidase activity and melibiose transport activity, which were induced by melibiose. Neither Na + nor Li + stimulated the melibiose transport. However, transport of methyl- β-thiogalactoside (TMG) was stimulated by Li + but not by Na +. These findings suggest that the major coupling cation for the melibiose transporter in E. aerogenes is H +. In fact, we observed H + entry into cells caused by an influx of melibiose and some of its analogs. We cloned the melB gene which encodes the melibiose transporter, and sequenced it. Deduced amino acid sequence of the transporter revealed that the melibiose transporter consists of 471 amino acid residues and the molecular weight was calculated to be 52 214 Da. The sequence showed high homology with the sequences of the melibiose transporters of Escherichia coli, Salmonella typhimurium and Klebsiella pneumoniae. Higher homology was found with the melibiose transporter of K. pneumoniae than with that of E. coli and S. typhimurium.