Phosphoenolpyruvate-dependent Maltose:phosphotransferase Activity in Fusobacterium Mortiferum ATCC 25557: Specificity, Inducibility, and Product Analysis

J Bacteriol. 1994 Jun;176(11):3250-6. doi: 10.1128/jb.176.11.3250-3256.1994.

Abstract

Phosphoenolypyruvate-dependent maltose:phosphotransferase activity was induced in cells of Fusobacterium mortiferum ATCC 25557 during growth on maltose. The disaccharide was rapidly metabolized by washed cells maintained under anaerobic conditions, but fermentation ceased immediately upon exposure of the cell suspension to air. Coincidentally, high levels of a phosphorylated derivative accumulated within the cells. Chemical and enzymatic analyses, in conjunction with data from 1H, 13C, and 31P nuclear magnetic resonance spectroscopy, established the structure of the purified compound as 6-O-phosphoryl-alpha-D-glucopyranosyl-(1-4)-D-glucose (maltose 6-phosphate). A method for the preparation of substrate amounts of this commercially unavailable disaccharide phosphate is described. Permeabilized cells of F. mortiferum catalyzed the phosphoenolpyruvate-dependent phosphorylation of maltose under aerobic conditions. However, the hydrolysis of maltose 6-phosphate (to glucose 6-phosphate and glucose) by permeabilized cells or cell-free preparations required either an anaerobic environment or addition of dithiothreitol to aerobic reaction mixtures. The first step in dissimilation of the phosphorylated disaccharide appears to be catalyzed by an oxygen-sensitive maltose 6-phosphate hydrolase. Cells of F. mortiferum, grown previously on maltose, fermented a variety of alpha-linked glucosides, including maltose, turanose, palatinose, maltitol, alpha-methylglucoside, trehalose, and isomaltose. Conversely, cells grown on the separate alpha-glucosides also metabolized maltose. For this anaerobic pathogen, we suggest that the maltose:phosphotransferase and maltose 6-phosphate hydrolase catalyze the phosphorylative translocation and cleavage not only of maltose but also of structurally analogous alpha-linked glucosides.

MeSH terms

  • Aerobiosis
  • Anaerobiosis
  • Carbohydrate Sequence
  • Cell Membrane Permeability
  • Enzyme Induction
  • Fusobacterium / enzymology*
  • Glucosides / metabolism
  • Hydrolysis
  • Maltose / metabolism*
  • Molecular Sequence Data
  • Phosphoenolpyruvate Sugar Phosphotransferase System / metabolism*
  • Phosphorylation
  • Stereoisomerism
  • Sugar Phosphates / isolation & purification*
  • Sugar Phosphates / metabolism

Substances

  • Glucosides
  • Sugar Phosphates
  • maltose 6-phosphate
  • Maltose
  • Phosphoenolpyruvate Sugar Phosphotransferase System