Exploring the sequence diversity in glycoside hydrolase family 13_18 reveals a novel glucosylglycerol phosphorylase

Appl Microbiol Biotechnol. 2018 Apr;102(7):3183-3191. doi: 10.1007/s00253-018-8856-1. Epub 2018 Feb 22.


In the carbohydrate-active enzyme database, GH13_18 is a family of retaining glycoside phosphorylases that act on α-glucosides. In this work, we explored the functional diversity of this family by comparing distinctive sequence motifs in different branches of its phylogenetic tree. A glycoside phosphorylase from Marinobacter adhaerens HP15 that was predicted to have a novel function was expressed and characterised. The enzyme was found to catalyse the reversible phosphorolysis of 2-O-α-D-glucosylglycerol with retention of the anomeric configuration, a specificity that has never been described before. Homology modelling, docking and mutagenesis were performed to pinpoint particular acceptor site residues (Tyr194, Ala333, Gln336) involved in the binding of glycerol. The new enzyme specificity provides additional insights into bacterial metabolic routes, being the first report of a phosphorolytic route for glucosylglycerol in a glucosylglycerol-producing organism. Furthermore, glucosylglycerol phosphorylase might be an attractive biocatalyst for the production of the osmolyte glucosylglycerol, which is currently produced on industrial scale by exploiting a side activity of the closely related sucrose phosphorylase. Family GH13_18 has clearly proven to be more diverse than was initially assumed, and the analysis of specificity-determining sequence motifs has shown to be a straightforward and fruitful tool for enzyme discovery.

Keywords: Glucosylglycerol; Glucosylglycerol phosphorylase; Glycoside hydrolase family GH13; Sucrose phosphorylase.

MeSH terms

  • Amino Acid Motifs
  • Bacterial Proteins / chemistry
  • Bacterial Proteins / genetics
  • Bacterial Proteins / metabolism
  • Genetic Variation*
  • Glucosides / metabolism
  • Glycoside Hydrolases / chemistry
  • Glycoside Hydrolases / genetics*
  • Glycoside Hydrolases / metabolism*
  • Marinobacter / enzymology*
  • Marinobacter / genetics*
  • Phosphorylases / chemistry
  • Phosphorylases / genetics*
  • Substrate Specificity


  • Bacterial Proteins
  • Glucosides
  • glucosylglycerol
  • Phosphorylases
  • Glycoside Hydrolases