Any feedback?
Please rate this page
(literature.php)
(0/150)

BRENDA support

Literature summary for 2.4.1.7 extracted from

  • Wiesbauer, J.; Goedl, C.; Schwarz, A.; Brecker, L.; Nidetzky, B.
    Substitution of the catalytic acid-base Glu237 by Gln suppresses hydrolysis during glucosylation of phenolic acceptors catalyzed by Leuconostoc mesenteroides sucrose phosphorylase (2010), J. Mol. Catal. B, 65, 24-29.
No PubMed abstract available
Search for more literature for 2.4.1.7

Cloned(Commentary)

Cloned (Comment) Organism
expression of His-tagged wild-type and mutant enzymes in Escherichia coli strain DH10B Leuconostoc mesenteroides

Protein Variants

Protein Variants Comment Organism
E237Q site-directed mutagenesis, replacement of the catalytic acid-base Glu237, the mutant does not display hydrolase activity under transglucosylation conditions and therefore provides 7fold enhancement of transfer yield Leuconostoc mesenteroides

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
additional information
-
 additional information kinetic mechanism for transglucosylation to external acceptors catalyzed by sucrose phosphorylase under conditions in which the natural acceptor substrate phosphate is absent, overview Leuconostoc mesenteroides

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
sucrose + phosphate Leuconostoc mesenteroides
-
 D-fructose + alpha-D-glucose 1-phosphate
-
 r

Organism

Organism UniProt Comment Textmining
Leuconostoc mesenteroides
-
-
-

Purification (Commentary)

Purification (Comment) Organism
recombinant His-tagged wild-type and mutant enzyme from Escherichia coli strain DH10B by nickel affinity chromatography Leuconostoc mesenteroides

Reaction

Reaction Comment Organism Reaction ID
sucrose + phosphate = D-fructose + alpha-D-glucose 1-phosphate catalytic mechanism of sucrose phosphorylase utilized for phosphorolysis of sucrose, hydrolysis, and transfer to acceptors. The requirement for base catalytic facilitation by Glu237 during deglucosylation of the enzyme will depend on the acceptor used, overview Leuconostoc mesenteroides
a

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
additional information both wild-type and mutated enzyme employ 4-nitrophenyl-alpha-D-glucopyranoside as a slow artificial substrate for phosphorolysis and hydrolysis Leuconostoc mesenteroides ?
-
 ?
sucrose + 2,6-difluorophenol with the wild-type enzyme, hydrolysis of the sugar 1-phosphate prevails about 10fold over glucosyl transfer to the 2,6-difluorophenol acceptor. Glucosylation of 2,6-difluorophenol is also catalyzed by enzyme mutant E237Q Leuconostoc mesenteroides D-fructose + 2,6-difluorophenyl alpha-D-glucoside
-
 r
sucrose + phosphate
-
 Leuconostoc mesenteroides D-fructose + alpha-D-glucose 1-phosphate
-
 r

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
30
-
 assay at Leuconostoc mesenteroides

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
7
-
 assay at Leuconostoc mesenteroides

General Information

General Information Comment Organism
malfunction in a series of mono- and disubstituted phenols differing in hydroxyl pKa between 7.02 and 8.71, the transferase activity of E237Q is dependent on steric rather than electronic properties of the acceptor used. The mutant does not display hydrolase activity under transglucosylation conditions and therefore provides 7fold enhancement of transfer yield. Structure-activity relationship analysis for glucosyl transfer to phenolic acceptors by E237Q, overview Leuconostoc mesenteroides