Structure-based enhancement of the first anomeric glucokinase

Promoting promiscuity. The availability of D-glucose-1-phosphate and its derivatives directly contributes to the efficiency of in vitro and in vivo glycosylation methodologies, such as natural-product glycorandomization. While one-step enzymatic routes to this vital class of sugar phosphates provide an attractive alternative to multistep chemical syntheses, no sugar kinase studied thus far is known to anomerically phosphorylate D-glucose. Herein we report both the discovery of inherent glucokinase (GlcK) activity in wild-type Lactococcus lactis galactokinase and the enhancement of this first GlcK to provide a catalyst capable of accepting a wide range of new monosaccharide substrates.