Nature and properties of hexitol transport systems in Escherichia coli

Abstract

In E. coli K 12 the naturally occurring hexitols D mannitol, D glucitol, and galactitol are taken up and phosphorylated via three distinct transport systems by a mechanism called either group translocation or vectorial phosphorylation. For every system, a membrane bound enzyme II complex of the phosphoenolpyruvate dependent phosphotransferase system has been found, each requiring phosphoenolpyruvate, enzyme I, and HPr or alternatively P HPr as the phosphate donor. Cells with a constitutive synthesis of all hexitol transport systems but with low P HPr levels have very low transport and phosphorylating activities in vivo, although 40 to 90% of the enzyme II complex activities are detected in cell extracts of such mutants. No indications for additional hexitol transport systems, especially for systems able to transport and accumulate free hexitols as in Klebsiella aerogenes, have been found. Substrate specificity, Km, and V(max) of the three transport systems for several hexitols and hexitol analogues have been determined by growth rates, transport activities, and in vitro phosphorylating activities. Each system was found to take up several hexitols, but only one hexitol serves as the inducer. This inducer invariably is the substrate with the highest affinity. Since bacterial transport systems, as a general rule, seem to have a relatively broad substrate specificity, in contrast to a more restricted inducer specificity, it is proposed to name the system inducible by D mannitol and coded by the gene mtlA the D mannitol transport system, the system inducible by D glucitol and coded by gutA the D glucitol transport system, and the system inducible by galactitol and coded by gatA the galactitol transport system.