Abstract
An enzymic method is presented for the preparation of 14C-labelled and C1-deuterated D-glucose, to be used as a radioactive tracer in metabolic studies dealing with the fate of D-[1-2H]glucose. The procedure takes advantage of the deuteration of 14C-labelled D-fructose 6-phosphate during exposure to phosphomannoisomerase in D2O. It further includes the conversion of the deuterated hexose monophosphates to D-fructose 1,6-bisphosphate, its isolation and hydrolysis to D-fructose 6-phosphate and the partial conversion of the latter ester to D-glucose 6-phosphate, which is then isolated by ion exchange chromatography and hydrolyzed to unesterified D-glucose. The extent of deuteration on the C1 of D-glucose was judged, after conversion to D-glucose 6-phosphate, from the isotopic discrimination in reaction velocity exerted towards the C1-deuterated ester by glucose-6-phosphate dehydrogenase. The degree of isotopic discrimination was virtually identical to that found with D-glucose 6-phosphate prepared from pure D-[1-2H]glucose.
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