Protein Variants | Comment | Organism |
---|---|---|
D295E | mutation decreases the catalytic center activity of sucrose phosphorylase to about 0.01% of the wild-type level. The 100000fold preference of the wild-type for glucosyl transfer compared with mannosyl transfer from phosphate to fructose is lost | Leuconostoc mesenteroides |
D295E | site-directed mutagenesis of the catalytic residue, the mutant shows about 0.01% of the wild-type enzyme activity, the preference of the wild-type enzyme for glucosyl transfer compared with mannosyl transfer from phosphate to fructose is lost in the mutant enzyme | Leuconostoc mesenteroides |
D295N | mutation decreases the catalytic center activity of sucrose phosphorylase to about 0.01% of the wild-type level. Glucosylation and deglucosylation steps are affected uniformly, and independently of leaving group ability and nucleophilic reactivity of the substrate, respectively. The 100000fold preference of the wild-type for glucosyl transfer compared with mannosyl transfer from phosphate to fructose is lost | Leuconostoc mesenteroides |
D295N | site-directed mutagenesis of the catalytic residue, the mutant shows about 0.01% of the wild-type enzyme activity, the preference of the wild-type enzyme for glucosyl transfer compared with mannosyl transfer from phosphate to fructose is lost in the mutant 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 |
---|---|
mutant enzymes D295N and D295E | Leuconostoc mesenteroides |
Reaction | Comment | Organism | Reaction ID |
---|---|---|---|
sucrose + phosphate = D-fructose + alpha-D-glucose 1-phosphate | reaction mechanism, the side chain of Asp295, through a strong hydrogen bond with the equatorial sugar 2-hydroxyl, stabilizes the transition states flanking the bglucosyl enzyme intermediate | Leuconostoc mesenteroides |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
alpha-D-glucose 1-phosphate + arsenate | - |
Leuconostoc mesenteroides | ? | - |
? | |
additional information | glucosyl donor and acceptor specificities, in absence of acceptor, the enzyme performs hydrolysis of alpha-D-glucose 1-phosphate | Leuconostoc mesenteroides | ? | - |
? | |
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 |
---|---|---|---|---|---|
additional information | - |
additional information | kinetics of recombinant wild-type and mutant enzymes, overview | Leuconostoc mesenteroides | |
0.0014 | - |
D-fructose | pH 7.0, 30°C, synthesis, mutant D295E | Leuconostoc mesenteroides | |
0.0065 | - |
arsenate | pH 7.0, 30°C, arsenolysis, mutant D295E | Leuconostoc mesenteroides | |
0.0124 | - |
alpha-D-glucose 1-phosphate | pH 7.0, 30°C, synthesis, mutant D295E | Leuconostoc mesenteroides | |
0.0136 | - |
phosphate | pH 7.0, 30°C, phosphorolysis, mutant D295E | Leuconostoc mesenteroides | |
0.0159 | - |
alpha-D-glucose 1-phosphate | pH 7.0, 30°C, synthesis, mutant D295N | Leuconostoc mesenteroides | |
0.02 | - |
sucrose | pH 7.0, 30°C, phosphorolysis, mutant D295N | Leuconostoc mesenteroides | |
0.0206 | - |
D-fructose | pH 7.0, 30°C, synthesis, mutant D295N | Leuconostoc mesenteroides | |
0.024 | - |
phosphate | pH 7.0, 30°C, phosphorolysis, mutant D295N | Leuconostoc mesenteroides | |
0.0252 | - |
arsenate | pH 7.0, 30°C, arsenolysis, mutant D295N | Leuconostoc mesenteroides | |
1.4 | - |
D-fructose | 30°C, pH 7.0, mutant enzyme D295E | Leuconostoc mesenteroides | |
1.59 | - |
alpha-D-glucose 1-phosphate | 30°C, pH 7.0, mutant enzyme D295N | Leuconostoc mesenteroides | |
1.99 | - |
sucrose | 30°C, pH 7.0, mutant enzyme D295N | Leuconostoc mesenteroides | |
2.06 | - |
D-fructose | 30°C, pH 7.0, mutant enzyme D295N | Leuconostoc mesenteroides | |
2.38 | - |
phosphate | 30°C, pH 7.0, mutant enzyme D295N | Leuconostoc mesenteroides | |
12.4 | - |
alpha-D-glucose 1-phosphate | 30°C, pH 7.0, mutant enzyme D295E | Leuconostoc mesenteroides | |
13.6 | - |
phosphate | 30°C, pH 7.0, mutant enzyme D295E | Leuconostoc mesenteroides | |
72 | - |
D-fructose | 30°C, pH 7.0, wild-type enzyme | Leuconostoc mesenteroides | |
72 | - |
D-fructose | pH 7.0, 30°C, synthesis, wild-type enzyme | Leuconostoc mesenteroides | |
91 | - |
alpha-D-glucose 1-phosphate | 30°C, pH 7.0, wild-type enzyme | Leuconostoc mesenteroides | |
91 | - |
alpha-D-glucose 1-phosphate | pH 7.0, 30°C, synthesis, wild-type enzyme | Leuconostoc mesenteroides | |
165 | - |
sucrose | 30°C, pH 7.0, wild-type enzyme | Leuconostoc mesenteroides | |
165 | - |
sucrose | pH 7.0, 30°C, phosphorolysis, wild-type enzyme | Leuconostoc mesenteroides | |
170 | - |
arsenate | pH 7.0, 30°C, arsenolysis, wild-type enzyme | Leuconostoc mesenteroides | |
195 | - |
phosphate | 30°C, pH 7.0, wild-type enzyme | Leuconostoc mesenteroides | |
195 | - |
phosphate | pH 7.0, 30°C, phosphorolysis, wild-type enzyme | Leuconostoc mesenteroides | |
261 | - |
alpha-D-glucose 1-phosphate | pH 7.0, 30°C, arsenolysis, wild-type enzyme | Leuconostoc mesenteroides |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
7 | - |
assay at | Leuconostoc mesenteroides |
pH Minimum | pH Maximum | Comment | Organism |
---|---|---|---|
additional information | - |
pH-dependence of wild-type and mutant enzymes, overview | Leuconostoc mesenteroides |