An enzymatic synthesis of the glycolytic intermediate,2-phospho-D-glycerate,is reported.This synthesis was undertaken as a means to make 2-phosho-D-glycerate quickly and inexpensively,because this compound is no longer available commercially.The synthesis uses the recombinant enzyme,D-glycerate-2-kinase,and the starting compounds adenosine 5'-triphosphate and D-glycerate from racemic D-L-glycerate.This enzymatic reaction is both stereospecific and regiospecific,producing 2-phospho-D-glycerate exclusively.The product is purified by anion-exchange chromatography using gradient elution with a volatile buffer.This straightforward method produces the potassium salt of 2-phosho-D-glycerate in high yield.The expression and purification of D-glycerate-2-kinase also are discussed.
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