Albermann Christoph, Weiner Michael, Tröndle Julia, Weuster-Botz Dirk, Sprenger Georg A
Institute of Microbiology, Universität Stuttgart, Stuttgart, Germany
Institute of Biochemical Engineering, Technische Universität München, Garching, Germany.
FEMS Microbiol Lett. 2014 Dec;361(1):52-61. doi: 10.1111/1574-6968.12612. Epub 2015 Jan 8.
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-labeling experiments.
磷酸盐反向转运蛋白介导。在本研究中，我们展示了重组磷酸烯醇丙酮酸：磷酸盐反向转运蛋白在大肠杆菌菌株同位素标记实验中的应用。研究了反向转运蛋白UhpT、UhpT-D388C和PgtP在磷酸烯醇丙酮酸作为唯一碳源时的转运活性和生长情况。在产莽草酸的大肠杆菌菌株中表达蛋白变体UhpT-D388C，以显示从细胞外磷酸烯醇丙酮酸成功对莽草酸进行同位素标记。结果证明了将外源应用的糖酵解中间产物直接掺入大肠杆菌细胞用于C标记实验的可能性。
