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Characterization of Membrane Transporters by Heterologous Expression in E. coli and Production of Membrane Vesicles
Published on: December 31, 2019
Characterization of the dicarboxylate transporter DctA in Corynebacterium glutamicum
Jung-Won Youn1, Elena Jolkver, Reinhard Krämer
1Institute of Molecular Microbiology and Biotechnology, Westfalian Wilhelms University Muenster, D-48149 Muenster, Germany.
Abstract:
Transporters of the dicarboxylate amino acid-cation symporter family often mediate uptake of C(4)-dicarboxylates, such as succinate or l-malate, in bacteria. A member of this family, dicarboxylate transporter A (DctA) from Corynebacterium glutamicum, was characterized to catalyze uptake of the C(4)-dicarboxylates succinate, fumarate, and l-malate, which was inhibited by oxaloacetate, 2-oxoglutarate, and glyoxylate. DctA activity was not affected by sodium availability but was dependent on the electrochemical proton potential. Efficient growth of C. glutamicum in minimal medium with succinate, fumarate, or l-malate as the sole carbon source required high dctA expression levels due either to a promoter-up mutation identified in a spontaneous mutant or to ectopic overexpression. Mutant analysis indicated that DctA and DccT, a C(4)-dicarboxylate divalent anion/sodium symporter-type transporter, are the only transporters for succinate, fumarate, and l-malate in C. glutamicum.
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