Biocatalytic Asymmetric Phosphorylation Catalyzed by Recombinant Glycerate‐2‐Kinase. (19th June 2017) - Hardt, Norman
Abstract: The efficient synthesis of pured ‐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 31 P NMR and showed excellent enantioselectivity towards phosphorylation of thed ‐enantiomer of glyceric acid. This straightforward phosphorylation reaction and subsequent product isolation enabled the preparation of enantiomerically pured ‐glycerate 2‐phosphate. This phosphorylation reaction, using recombinant glycerate‐2‐kinase, yieldedd ‐glycerate 2‐phosphate in fewer reaction steps and with higher purity than chemical routes. Abstract : Quicker and cleaner : Recombinant glycerate‐2‐kinase from T. maritima showed excellent selectivity towards phosphorylation of thed ‐enantiomer of glyceric acid, with no phosphorylation of thel ‐enantiomer. This straightforward phosphorylation reaction and subsequent product isolation enables the central glycolytic metabolited ‐glycerate 2‐phosphate to be prepared in fewer reaction steps and with higher purity than chemical routes.