Utilization of organophosphate:phosphate antiporter for isotope‐labeling experiments in E. coli
2014
Albermann, Christoph | Weiner, Michael | Tröndle, Julia | Weuster‐Botz, Dirk | Sprenger, Georg A.
The transport of organophosphates across the cytoplasma membrane is mediated by organophosphate:phosphate antiporter proteins. In this work, we present the application of a recombinant phosphoenolpyruvate:phosphate antiporter for isotopic labeling experiments in E. coli strains. The antiporters UhpT, UhpT‐D388C, and PgtP were investigated regarding transport activity and growth on phosphoenolpyruvate as sole carbon source. The expression of the protein variant UhpT‐D388C in a shikimic acid producing E. coli strain was used to show the successful isotopic labeling of shikimic acid from extracellular phosphoenolpyruvate. The results demonstrate the possibility of a direct incorporation of exogenously applicated glycolysis intermediates into E. coli cells for¹³C‐labeling experiments.
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