Structural Characterization of Linear isomalto-/malto-oligomer Products Synthesized by the Novel GTFB 4,6-α-glucanotransferase Enzyme From Lactobacillus Reuteri 121

Glycobiology. 2012 Apr;22(4):517-28. doi: 10.1093/glycob/cwr167. Epub 2011 Dec 2.

Abstract

Recently, a novel glucansucrase (GS)-like gene (gtfB) was isolated from the probiotic bacterium Lactobacillus reuteri 121 and expressed in Escherichia coli. The purified recombinant GTFB enzyme was characterized and turned out to be inactive with sucrose, the natural GS substrate. Instead, GTFB acted on malto-oligosaccharides (MOSs), thereby yielding elongated gluco-oligomers/polymers containing besides (α1 → 4) also (α1 → 6) glycosidic linkages, and it was classified as a 4,6-α-glucanotransferase. To gain more insight into its reaction specificity, incubations of the GTFB enzyme with a series of MOSs and their corresponding alditols [degree of polymerization, DP2(-ol)-DP7(-ol)] were carried out, and (purified) products were structurally analyzed with matrix-assisted laser desorption ionization time-of-flight mass spectrometry and one-/two-dimensional (1)H and (13)C nuclear magnetic resonance spectroscopy. With each of the tested malto-oligomers, the GTFB enzyme yielded series of novel linear isomalto-/malto-oligomers, in the case of DP7 up to DP >35.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Bacterial Proteins / chemistry*
  • Carbohydrate Conformation
  • Carbohydrate Sequence
  • Chromatography, Ion Exchange
  • Glucans / chemistry
  • Glucosyltransferases / chemistry*
  • Glycosylation
  • Lactobacillus reuteri / enzymology*
  • Magnetic Resonance Spectroscopy
  • Maltose / chemistry*
  • Molecular Sequence Data
  • Oligosaccharides / chemical synthesis*
  • Oligosaccharides / chemistry
  • Oligosaccharides / isolation & purification
  • Recombinant Proteins / chemistry
  • Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
  • Sugar Alcohols / chemistry
  • Trisaccharides / chemistry

Substances

  • Bacterial Proteins
  • Glucans
  • Oligosaccharides
  • Recombinant Proteins
  • Sugar Alcohols
  • Trisaccharides
  • maltoheptaose
  • maltotriose
  • Maltose
  • Glucosyltransferases