27] Thiamine pyrophosphate-agarose and the purification of a thiamine-binding protein

Abstract

Publisher Summary Escherichia coli contains a thiamine-binding protein that is thought to be involved in the transport of thiamine through the cell membrane. Because of the very small amount of the thiamine-binding protein in the periplasmic space of the cells, a more effective purification procedure than the conventional method was required to isolate reasonable amounts of the highly purified protein. During an investigation into the properties of the thiamine-binding protein from E. coli, it has been found that the partially purified protein can bind to thiamine pyrophosphate (TPP) and thiamine monophosphate (TMP), as well as to thiamine. This finding suggested that coupling of the thiamine phosphates to an insoluble matrix might yield an efficient affinity chromatographic system.