Sodium-Dependent Uptake of Glutamate by Novel ApGltS Enhanced Growth under Salt Stress of Halotolerant Cyanobacterium Aphanothece halophytica

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  • BOONBURAPONG Bongkoj
    Graduate School of Environmental and Human Sciences, Meijo University Faculty of Science, Chulalongkorn University Graduate School of Environmental and Human Sciences, Meijo University Faculty of Science, Chulalongkorn University
  • LALOKNAM Surasak
    Research Institute of Meijo University Faculty of Science, Srinakharinwirot University Research Institute of Meijo University Faculty of Science, Srinakharinwirot University
  • YAMADA Nana
    Graduate School of Environmental and Human Sciences, Meijo University Graduate School of Environmental and Human Sciences, Meijo University
  • INCHAROENSAKDI Aran
    Faculty of Science, Chulalongkorn University Faculty of Science, Chulalongkorn University
  • TAKABE Teruhiro
    Graduate School of Environmental and Human Sciences, Meijo University Research Institute of Meijo University Graduate School of Environmental and Human Sciences, Meijo University Research Institute of Meijo University

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  • Sodium-Dependent Uptake of Glutamate by Novel ApGltS Enhanced Growth under Salt Stress of Halotolerant Cyanobacterium <i>Aphanothece halophytica</i>

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Glutamate is a major free amino acid in cyanobacteria, but its transport properties remain largely unknown. In this study, we found that a halotolerant cyanobacterium, Aphanothece halophytica, contained a sodium dependent glutamate transporter (ApGltS). The deduced amino acid sequence of ApGltS exhibited low homology (18–19% identity) to GltS from Synechocystis sp. PCC 6803 (slr1145) and Escherichia coli. The predicted ApGltS consisted of 476 amino acid residues with a molecular weight of 50,976 Da. As analysed by hydropathy profiling, ApGltS contains 11 transmembrane segments. The ApgltS gene was isolated and expressed in E. coli ME9107, which is deficient in glutamate uptake. ME9107, expressing ApGltS, took up glutamate and its rates increased with increasing concentrations of NaCl. Kinetics studies revealed that ApGltS is a high-affinity glutamate transporter with a Km of about 5 µM. The presence of 0.5 M NaCl in the assay medium increased Vmax by about 3-fold. Competition experiments revealed that glutamate, glutamine, aspartate, and asparagine inhibited glutamate uptake. The level of mRNA for ApgltS was higher in A. halophytica grown at high salinity. Under high salinity conditions supplemented with glutamate, A. halophytica showed a significant increase in intracellular glycine betaine.

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