Trehalose-6-phosphate phosphorylase is part of a novel metabolic pathway for trehalose utilization in Lactococcus lactis

J Biol Chem. 2001 Nov 16;276(46):42707-13. doi: 10.1074/jbc.M108279200. Epub 2001 Sep 11.


Lactococcus lactis splits phosphorylated trehalose by the action of inorganic phosphate-dependent trehalose-6-phosphate phosphorylase (TrePP) in a novel catabolic pathway. TrePP was found to catalyze the reversible conversion of trehalose 6-phosphate into beta-glucose 1-phosphate and glucose 6-phosphate by measuring intermediate sugar phosphates in cell extracts from trehalose-cultivated lactococci. According to native PAGE and SDS-PAGE, TrePP was shown to be a monomeric enzyme with a molecular mass of 94 kDa. Reaction kinetics suggested that the enzyme follows a ternary complex mechanism with optimal phosphorolysis at 35 degrees C and pH 6.3. The equilibrium constants were found to be 0.026 and 0.032 at pH 6.3 and 7.0, respectively, favoring the formation of trehalose 6-phosphate. The Michaelis-Menten constants of TrePP for trehalose 6-phosphate, inorganic phosphate, beta-glucose 1-phosphate, and glucose 6-phosphate were determined to be 6, 32, 0.9, and 4 mm, respectively. The TrePP-encoding gene, designated trePP, was localized in a putative trehalose operon of L. lactis. This operon includes the gene encoding beta-phosphoglucomutase in addition to three open reading frames believed to encode a transcriptional regulator and two trehalose-specific phosphotransferase system components. The identity of trePP was confirmed by determining the N-terminal amino acid sequence of TrePP and by its overexpression in Escherichia coli and L. lactis, as well as the construction of a lactococcal trePP knockout mutant. Furthermore, both TrePP and beta-phosphoglucomutase activity were detected in Enterococcus faecalis cell extract, indicating that this bacterium exhibits the same trehalose assimilation route as L. lactis.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acids / chemistry
  • Blotting, Southern
  • Electrophoresis, Polyacrylamide Gel
  • Enterococcus faecalis / enzymology
  • Escherichia coli / metabolism
  • Glucose-6-Phosphate / metabolism
  • Glucosephosphates / metabolism
  • Glucosyltransferases / chemistry*
  • Glucosyltransferases / isolation & purification
  • Glucosyltransferases / metabolism*
  • Glucosyltransferases / physiology*
  • Hydrogen-Ion Concentration
  • Kinetics
  • Lactococcus lactis / enzymology*
  • Lactococcus lactis / metabolism*
  • Models, Biological
  • Phosphoglucomutase / chemistry
  • Phosphoglucomutase / genetics
  • Phylogeny
  • Plasmids / metabolism
  • Protein Binding
  • Stereoisomerism
  • Temperature
  • Time Factors
  • Transcription, Genetic
  • Trehalose / metabolism*


  • Amino Acids
  • Glucosephosphates
  • Glucose-6-Phosphate
  • Trehalose
  • glucose-1-phosphate
  • Glucosyltransferases
  • trehalose-6-phosphate phosphorylase
  • alpha,alpha-trehalose phosphorylase
  • Phosphoglucomutase