Cloned (Comment) | Organism |
---|---|
expression of His-tagged wild-type and mutant enzymes in Escherichia coli strain DH10B | Leuconostoc mesenteroides |
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 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 | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
sucrose + phosphate | Leuconostoc mesenteroides | - |
D-fructose + alpha-D-glucose 1-phosphate | - |
r |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Leuconostoc mesenteroides | - |
- |
- |
Purification (Comment) | Organism |
---|---|
recombinant His-tagged wild-type and mutant enzyme from Escherichia coli strain DH10B by nickel affinity chromatography | Leuconostoc mesenteroides |
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 |
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 Maximum [°C] | Comment | Organism |
---|---|---|---|
30 | - |
assay at | Leuconostoc mesenteroides |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
7 | - |
assay at | Leuconostoc mesenteroides |
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 |