Biocatalytic Asymmetric Phosphorylation Catalyzed by Recombinant Glycerate-2-Kinase.

The efficient synthesis of pure D-glycerate-2-phosphate is of great interest due to its importance as an enzyme substrate and metabolite. Therefore, we investigated a straightforward one-step biocatalytic phosphorylation of glyceric acid. Glycerate-2-kinase from Thermotoga maritima was expressed in Escherichia coli, allowing easy purification. The selective glycerate-2-kinase-catalyzed phosphorylation was followed by P-31 NMR and showed excellent enantioselectivity towards phosphorylation of the D-enantiomer of glyceric acid. This straightforward phosphorylation reaction and subsequent product isolation enabled the preparation of enantiomerically pure D-glycerate 2-phosphate. This phosphorylation reaction, using recombinant glycerate-2-kinase, yielded D-glycerate 2-phosphate in fewer reaction steps and with higher purity than chemical routes.