An Escherichia coli mutant defective in high-affinity D-ribose transport is able to utilize the sugar as a sole carbon source, suggesting that other transport systems for D-ribose exist. In order to search for such transporters, transposon mutagenesis was carried out in an rbsB-negative strain containing ribokinase (rbsK) for sugar phosphorylation. Insertions showing an enhanced ribose growth were isolated and mapped in xylA and its promoter region. The mutations increased not only the ribose uptake but also the expression of xylFGH encoding an ABC (ATP-binding cassette)-type transporter for D-xylose. Secondary mutations abolishing the ribose-utilizing phenotype were obtained both in the xylFG genes coding for the xylose high-affinity transporter and in xylR that is required for the xyl gene expression. Ribose uptake was also reduced by the secondary mutations. An overexpression of sylFGH under Pt-rc promoter supported enhanced growth on ribose. These results indicate that D-ribose can be transported through the XylFGH transporter. (C) 1998 Federation of European Microbiological Societies. Published by Elsevier Science B.V. All rights reserved.