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Characterization of Membrane Transporters by Heterologous Expression in E. coli and Production of Membrane Vesicles
Published on: December 31, 2019
Identification and expression analysis of a gene encoding a shikimate transporter of Corynebacterium glutamicum
Takeshi Kubota1, Yuya Tanaka1, Norihiko Takemoto1
1Research Institute of Innovative Technology for the Earth, 9-2 Kizugawadai, Kizugawa, Kyoto 619-0292, Japan.
Abstract:
Shikimate can be utilized as the sole source of carbon and energy of Corynebacterium glutamicum. Although biosynthesis and degradation of shikimate are well characterized in C. glutamicum, the transport of shikimate has hardly been studied. A mutant strain deficient in cgR_2523 loses the ability to grow on shikimate as well as to consume extracellular shikimate, indicating that the gene is involved in shikimate utilization (designated shiA). The hydropathy profile of the deduced amino acid sequence indicates that ShiA belongs to the metabolite/proton symporter family, which is a member of the major facilitator superfamily. An accumulation assay showed that the uptake of shikimate was hardly detected in the shiA-deficient strain, but was markedly enhanced in a shiA-expressing strain. These results suggested that the uptake of shikimate was mainly mediated by the shikimate transporter encoded by shiA. The level of shiA mRNA induction by shikimate was significantly decreased by the disruption of cgR_2524 (designated shiR), which is located immediately upstream of shiA and encodes a LysR-type transcriptional regulator, suggesting that ShiR acts as an activator of shiA. To our knowledge, this is the first report in Gram-positive bacteria of a shikimate transporter and its regulation.
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