Structural Basis for Reversible Phosphorolysis and Hydrolysis Reactions of 2-O-α-Glucosylglycerol Phosphorylase

Abstract

Background: 2-O-␣-Glucosylglycerol phosphorylase (GGP) catalyzes the reversible phosphorolysis of glucosylglycerol and the hydrolysis of ␤-D-glucose 1-phosphate. Results: Crystal structures were determined for GGP complexed with glucose, isofagomine, and glycerol. Conclusion: A water-mediated hydrogen bond network that fixes glycerol is the key factor of the bifunctionality. Significance: The substrate recognition and reaction mechanisms of GGP were clarified in detail.

Diversity of phosphorylases in glycoside hydrolase families

“…This is the only activity identified in the subfamily, and starch synthase is the only phosphorylase known to generate oligosaccharide 1-phosphate. The activity and cloning of the gene encoding this enzyme were first Nihira et al 2014c Touhara et al 2014 reported for Corynebacterium in a patent (Igarashi et al 2005). The same activity of Mycobacterium tuberculosis was reported in a journal article with an explanation of the role of this enzyme (Elbein et al 2010).…”

“…This enzyme possesses moderate hydrolytic activity on βGlc1P and weak phosphate-dependent hydrolytic activity on kojibiose. Structural analysis of the enzyme revealed the mechanism of hydrolysis of a sugar 1-phosphate, a unique activity among phosphorylases (Touhara et al 2014). The strain also possesses two other unique GH 65 enzymes, branching oligosaccharide-forming maltose phosphorylase (Nihira et al 2012d) and potassium ion-dependent inverting trehalose phosphorylase (Nihira et al 2013a).…”

Heterosides – compatible solutes occurring in prokaryotic and eukaryotic phototrophs

“…Only recently, has the first GG‐phosphorylase been identified and characterised from Bacillus selenitrireducens (Touhara et al . ).…”

“…Another GG synthesis reaction was described that reversibly phosphorylates GG to glucose-1-phosphate and glycerol, similar to the reversible reaction of sucrose phosphorylase. Only recently, has the first GG-phosphorylase been identified and characterised from Bacillus selenitrireducens (Touhara et al 2014).…”

Disaccharide phosphorylases: Structure, catalytic mechanisms and directed evolution

“…2 B). The domain organization of LbMP is similar to other GH65 disaccharide phosphorylases, which include kojibiose phosphorylase from Caldicellulosiruptor saccharolyticus (CsKP) [ 37 ] and glucosylglycerol phosphorylase from Bacillus selenitireducens (BsGGP) [ 39 ]. Moreover, Paenibacillus sp.…”