科研之友
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摘要
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 for C-13-labeling experiments.
