Cloned (Comment) | Organism |
---|---|
expression of Strep-tagged wild-type and mutant enzymes in Escherichia coli Top10 cells | Leuconostoc mesenteroides |
Protein Variants | Comment | Organism |
---|---|---|
F52A | site-directed mutagenesis, large destabilization of the transition states for enzyme glucosylation and deglucosylation in the mutant compared to the wild-type enzyme, while the relative stability of the glucosyl enzyme intermediate was weakly affected by substitution of Phe52 | Leuconostoc mesenteroides |
F52N | site-directed mutagenesis, large destabilization of the transition states for enzyme glucosylation and deglucosylation in the mutant compared to the wild-type enzyme, while the relative stability of the glucosyl enzyme intermediate was weakly affected by substitution of Phe52 | Leuconostoc mesenteroides |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
additional information | - |
additional information | free energy profiles for reactions of wild-type and mutated enzymes, and steady-state kinetic analysis, overview | Leuconostoc mesenteroides | |
4.7 | - |
alpha-D-glucose 1-phosphate | pH 7.0, 30°C, sucrose synthesis by glycosylation from alpha-D-glucose 1-phosphate, recombinant wild-type enzyme | Leuconostoc mesenteroides | |
6 | - |
sucrose | pH 7.0, 30°C, deglycosylation by phosphate, recombinant wild-type enzyme | Leuconostoc mesenteroides | |
9.8 | - |
sucrose | pH 7.0, 30°C, sucrose phosphorolysis, recombinant wild-type enzyme | Leuconostoc mesenteroides | |
13 | - |
sucrose | pH 7.0, 30°C, deglycosylation by D-fructose, recombinant wild-type enzyme | Leuconostoc mesenteroides | |
22 | - |
alpha-D-glucopyranosyl fluoride | pH 7.0, 30°C, phosphorolysis, recombinant wild-type enzyme | 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 Strep-tagged wild-type and mutant enzymes from Escherichia coli Top10 cells by single step affinity chromatography | Leuconostoc mesenteroides |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
alpha-D-glucopyranosyl fluoride + phosphate | - |
Leuconostoc mesenteroides | fluoride + alpha-D-glucose 1-phosphate | - |
? | |
D-fructose + alpha-D-glucose 1-phosphate | - |
Leuconostoc mesenteroides | sucrose + phosphate | - |
r | |
sucrose + phosphate | - |
Leuconostoc mesenteroides | D-fructose + alpha-D-glucose 1-phosphate | - |
r |
Synonyms | Comment | Organism |
---|---|---|
SPase | - |
Leuconostoc mesenteroides |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
30 | - |
assay at | Leuconostoc mesenteroides |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
39 | - |
alpha-D-glucose 1-phosphate | pH 7.0, 30°C, sucrose synthesis by glycosylation from alpha-D-glucose 1-phosphate, recombinant wild-type enzyme | Leuconostoc mesenteroides | |
47 | - |
sucrose | pH 7.0, 30°C, deglycosylation by D-fructose, recombinant wild-type enzyme | Leuconostoc mesenteroides | |
117 | - |
sucrose | pH 7.0, 30°C, sucrose phosphorolysis, recombinant wild-type enzyme | Leuconostoc mesenteroides | |
121 | - |
alpha-D-glucopyranosyl fluoride | pH 7.0, 30°C, phosphorolysis, recombinant wild-type enzyme | Leuconostoc mesenteroides | |
145 | - |
sucrose | pH 7.0, 30°C, deglycosylation by phosphate, recombinant wild-type enzyme | Leuconostoc mesenteroides |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
7 | - |
assay at | Leuconostoc mesenteroides |
General Information | Comment | Organism |
---|---|---|
evolution | the enzyme is a transglucosidase belonging to glycosylhydrolase family GH 13 | Leuconostoc mesenteroides |
kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
3.7 | - |
sucrose | pH 7.0, 30°C, deglycosylation by D-fructose, recombinant wild-type enzyme | Leuconostoc mesenteroides | |
5.5 | - |
alpha-D-glucopyranosyl fluoride | pH 7.0, 30°C, phosphorolysis, recombinant wild-type enzyme | Leuconostoc mesenteroides | |
8.3 | - |
alpha-D-glucose 1-phosphate | pH 7.0, 30°C, sucrose synthesis by glycosylation from alpha-D-glucose 1-phosphate, recombinant wild-type enzyme | Leuconostoc mesenteroides | |
12 | - |
sucrose | pH 7.0, 30°C, sucrose phosphorolysis, recombinant wild-type enzyme | Leuconostoc mesenteroides | |
24 | - |
sucrose | pH 7.0, 30°C, deglycosylation by phosphate, recombinant wild-type enzyme | Leuconostoc mesenteroides |