Structural analysis of rebaudioside A derivatives obtained by Lactobacillus reuteri 180 glucansucrase-catalyzed trans-α-glucosylation

Carbohydr Res. 2017 Feb 22:440-441:51-62. doi: 10.1016/j.carres.2017.01.008. Epub 2017 Jan 31.

Abstract

The wild-type Gtf180-ΔN glucansucrase enzyme from Lactobacillus reuteri 180 was found to catalyze the α-glucosylation of the steviol glycoside rebaudioside A, using sucrose as glucosyl donor in a transglucosylation process. Structural analysis of the formed products by MALDI-TOF mass spectrometry, methylation analysis and NMR spectroscopy showed that rebaudioside A is specifically α-d-glucosylated at the steviol C-19 β-d-glucosyl moiety (55% conversion). The main product is a mono-(α1 → 6)-glucosylated derivative (RebA-G1). A series of minor products, up to the incorporation of eight glucose residues, comprise elongations of RebA-G1 with mainly alternating (α1 → 3)- and (α1 → 6)-linked glucopyranose residues. These studies were carried out in the context of a program directed to the improvement of the taste of steviol glycosides via enzymatic modification of their naturally occurring carbohydrate moieties.

Keywords: Carbohydrate; Glycobiotechnology; NMR spectroscopy; Stevia rebaudiana; Steviol glycosides; Sweetener.

MeSH terms

  • Bacterial Proteins / chemistry*
  • Bacterial Proteins / isolation & purification
  • Carbohydrate Conformation
  • Catalysis
  • Diterpenes, Kaurane / chemistry*
  • Glucose / chemistry*
  • Glycosylation
  • Glycosyltransferases / chemistry*
  • Glycosyltransferases / isolation & purification
  • Limosilactobacillus reuteri / chemistry*
  • Limosilactobacillus reuteri / enzymology
  • Magnetic Resonance Spectroscopy
  • Methylation
  • Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
  • Stevia / chemistry
  • Sucrose / chemistry*

Substances

  • Bacterial Proteins
  • Diterpenes, Kaurane
  • steviol
  • Sucrose
  • rebaudioside A
  • Glycosyltransferases
  • alternansucrase
  • Glucose