Synthesis of FimH receptor-active manno-oligosaccharides by reverse hydrolysis using alpha-mannosidases from Penicillium citrinum, Aspergillus phoenicis and almond

Appl Microbiol Biotechnol. 2004 Feb;63(6):666-71. doi: 10.1007/s00253-003-1416-2. Epub 2003 Aug 9.

Abstract

Recombinant Penicillium citrinum alpha-1,2-mannosidase, expressed in Aspergillus oryzae, was employed to carry out regioselective synthesis of alpha- d-mannopyranosyl-(1-->2)- d-mannose. Yields (w/w) of 16.68% disaccharide, 3.07% trisaccharide and 0.48% tetrasaccharide were obtained, with alpha1-->2 linkages present at 98.5% of the total linkages formed. Non-specific alpha-mannosidase from almond was highly efficient in reverse hydrolysis and oligosaccharide yields of 45-50% were achieved. The products of the almond mannosidase were a mixture of disaccharides (30.75%, w/w), trisaccharides (12.26%, w/w) and tetrasaccharides (1.89%, w/w) with 1-->2, 1-->3 and 1-->6 isomers. alpha-1,2-linkage specific mannosidase from P. citrinum and alpha-1,6-linkage-specific mannosidase from Aspergillus phoenicis were used in combination to hydrolyse the respective linkages from the mixture of isomers, resulting in alpha- d-mannopyranosyl-(1-->3)- d-mannose in 86.4% purity. The synthesised oligosaccharides can potentially inhibit the adhesion of pathogens by acting as "decoys" of receptors of type-1 fimbriae carried by enterobacteria.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adhesins, Escherichia coli / metabolism
  • Aspergillus / enzymology
  • Aspergillus / genetics
  • Bacterial Adhesion / drug effects
  • Carbohydrate Conformation
  • Chromatography, Gel
  • Chromatography, High Pressure Liquid
  • Cloning, Molecular
  • Enzyme Stability
  • Fimbriae Proteins / metabolism
  • Genes, Fungal
  • Hydrogen-Ion Concentration
  • Hydrolysis
  • Mannosidases / metabolism
  • Mannosides / biosynthesis*
  • Oligosaccharides / analysis
  • Oligosaccharides / biosynthesis*
  • Oligosaccharides / chemistry
  • Oligosaccharides / isolation & purification*
  • Oligosaccharides / pharmacology
  • Penicillium / enzymology
  • Penicillium / genetics
  • Prunus / enzymology
  • Receptors, Immunologic / metabolism
  • Temperature
  • alpha-Mannosidase / genetics
  • alpha-Mannosidase / isolation & purification
  • alpha-Mannosidase / metabolism*

Substances

  • Adhesins, Escherichia coli
  • Mannosides
  • Oligosaccharides
  • Receptors, Immunologic
  • fimH protein, E coli
  • fimbriae receptor, type 1
  • Fimbriae Proteins
  • Mannosidases
  • mannosyl-oligosaccharide 1,2-alpha-mannosidase
  • alpha-Mannosidase