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Microbiology (2001), 147, 1069-1077.
© 2001 Society for General Microbiology

Genetics and Molecular Biology

The gene yghK linked to the glc operon of Escherichia coli encodes a permease for glycolate that is structurally and functionally similar to L-lactate permease

M. Felisa Núñez1, M. Teresa Pellicer1, Josefa Badía1, Juan Aguilar1 and Laura Baldomà1

Department of Biochemistry, Faculty of Pharmacy, University of Barcelona, Diagonal 643, 08028 Barcelona, Spain1

Author for correspondence: Juan Aguilar. Tel: +34 93 403 4496. Fax: +34 94 402 4520. e-mail: jaguilar{at}farmacia.far.ub.es

In Escherichia coli the glc operon involved in glycolate utilization is located at 67·3 min and formed by genes encoding the enzymes glycolate oxidase (glcDEF) and malate synthase G (glcB). Their expression from a single promoter upstream of glcD is induced by growth on glycolate and regulated by the activator encoded by the divergently transcribed gene glcC. Gene yghK, located 350 bp downstream of glcB, encodes a hydrophobic protein highly similar to the L-lactate permease encoded by lldP. Expression studies have shown that the yghK gene (proposed name glcA) is transcribed from the same promoter as the other glc structural genes and thus belongs to the glc operon. Characterization of a glcA::cat mutant showed that GlcA acts as glycolate permease and that glycolate can also enter the cell through another transport system. Evidence is presented of the involvement of L-lactate permease in glycolate uptake. Growth on this compound was abolished in a double mutant of the paralogous genes glcA and lldP, and restored with plasmids expressing either GlcA or LldP. Characterization of the putative substrates for these two related permeases showed, in both cases, specificity for the 2-hydroxymonocarboxylates glycolate, L-lactate and D-lactate. Although both GlcA and LldP recognize D-lactate, mutant analysis proved that L-lactate permease is mainly responsible for its uptake.

Keywords: gene function assignment, bacterial transport, LctP transporter family, paralogue genes

Abbreviations: CAA, casein acid hydrolysate; CAT, chloramphenicol acetyltransferase; LB, Luria–Bertani broth; IHF, integration host factor




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