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 alpha-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.
Kralj
Slavko
S
Centre for Carbohydrate Bioprocessing (CCB), TNO-University of Groningen, Haren, The Netherlands.
Eeuwema
Wieger
W
Eckhardt
Tom H
TH
Dijkhuizen
Lubbert
L
D001216:Asparagine / Q000502:physiology*; D001426:Bacterial Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D017124:Conserved Sequence; D004926:Escherichia coli / Q000235:genetics; D005960:Glucosides / Q000737:chemistry*; D006031:Glycosylation; D016695:Glycosyltransferases / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D007534:Isomaltose / Q000378:metabolism; D052200:Limosilactobacillus reuteri / Q000201:enzymology* / Q000235:genetics; D016297:Mutagenesis, Site-Directed; D009844:Oligosaccharides / Q000096:biosynthesis; D013057:Spectrum Analysis; D013379:Substrate Specificity; D013395:Sucrose / Q000378:metabolism
D001426:Bacterial Proteins; D005960:Glucosides; D009844:Oligosaccharides; D013395:Sucrose; D007534:Isomaltose; D001216:Asparagine; D016695:Glycosyltransferases; C033837:alternansucrase