Role of asparagine 1134 in glucosidic bond and transglycosylation specificity of reuteransucrase from Lactobacillus reuteri 121

Glucansucrases from lactic acid bacteria convert sucrose into various α‐glucans that differ greatly with respect to the glucosidic bonds present (e.g. dextran, mutan, alternan and reuteran). This study aimed to identify the structural features of the reuteransucrase from Lactobacillus reuteri 121 (G...

Full description

Saved in:
Bibliographic Details
Published in:The FEBS journal 2006-08, Vol.273 (16), p.3735-3742
Main Authors: Kralj, Slavko, Eeuwema, Wieger, Eckhardt, Tom H., Dijkhuizen, Lubbert
Format: Article
Language:English
Subjects:
Acids
Amino acids
Asparagine - physiology
Bacteria
Bacterial Proteins - chemistry
Bacterial Proteins - genetics
Bacterial Proteins - metabolism
Chemical bonds
Conserved Sequence
Enzymes
Escherichia coli - genetics
glucansucrase
Glucosides - chemistry
Glycosylation
Glycosyltransferases - chemistry
Glycosyltransferases - genetics
Glycosyltransferases - metabolism
Isomaltose - metabolism
Lactobacillus reuteri - enzymology
Lactobacillus reuteri - genetics
Lactobacillus reuteri
Mutagenesis, Site-Directed
Mutation
Oligosaccharides - biosynthesis
product specificity
reuteransucrase
site‐directed mutagenesis
Spectrum Analysis
Substrate Specificity
Sucrose - metabolism
Citations: Items that this one cites
Items that cite this one
Online Access:Get full text
Tags: Add Tag
No Tags, Be the first to tag this record!
Description
Summary:Glucansucrases from lactic acid bacteria convert sucrose into various α‐glucans that differ greatly with respect to the glucosidic bonds present (e.g. dextran, mutan, alternan and reuteran). This study aimed to identify the structural features of the reuteransucrase from Lactobacillus reuteri 121 (GTFA) that determine its reaction specificity. We here report a detailed mutational analysis of a conserved region immediately next to the catalytic Asp1133 (putative transition‐state stabilizing) residue in GTFA. The data show that Asn1134 is the main determinant of glucosidic bond product specificity in this reuteransucrase. Furthermore, mutations at this position greatly influenced the hydrolysis/transglycosylation ratio. Changes in this amino acid expands the range of glucan and gluco‐oligosaccharide products synthesized from sucrose by mutant GTFA enzymes.
ISSN:1742-464X
1742-4658
DOI:10.1111/j.1742-4658.2006.05376.x