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Literature summary for 2.4.1.7 extracted from
- Acid-base catalysis in Leuconostoc mesenteroides sucrose phosphorylase probed by site-directed mutagenesis and detailed kinetic comparison of wild-type and Glu237-Gln mutant enzymes (2007), Biochem. J., 403, 441-449.
Protein Variants
| Protein Variants | Comment | Organism |
|---|---|---|
| E237Q | site-directed mutagenesis of the catalytic residue, the mutant shows 0.001% of wild-type enzyme activity, reactions with substrates requiring Broensted catalytic assistance for glucosylation or deglucosylation are selectively slowed at the respective step about 10fold in mutant E237Q compared to the wild-type enzyme. Azide, acetate and formate but not halides restore catalytic activity up to 300fold in E237Q under conditions in which the deglucosylation step is rate-determining | Leuconostoc mesenteroides |
KM Value [mM]
| KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|
| additional information | - |
additional information | steady-state kinetics of wild-type and mutant E237Q enzymes | Leuconostoc mesenteroides |
Molecular Weight [Da]
| Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
|---|---|---|---|
| 55700 | - |
x * 55700, SDS-PAGE | 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 | - |
- |
- |
Reaction
| Reaction | Comment | Organism | Reaction ID |
|---|---|---|---|
| sucrose + phosphate = D-fructose + alpha-D-glucose 1-phosphate | reaction mechanism, acid-base catalysis in the two-step enzymatic mechanism of alpha-retaining glucosyl transfer by Leuconostoc mesenteroides sucrose phosphorylase, Glu237 is the catalytic acid-base of sucrose phosphorylase | Leuconostoc mesenteroides |
Specific Activity [micromol/min/mg]
| Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
|---|---|---|---|
| additional information | - |
- |
Leuconostoc mesenteroides |
| 0.0017 | - |
recombinant mutant E237Q | Leuconostoc mesenteroides |
Substrates and Products (Substrate)
| Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| 1-O-azido-alpha-D-glucose + phosphate | - |
Leuconostoc mesenteroides | ? | - |
? | |
| alpha-D-glucose 1-acetic acid ester + phosphate | alpha-D-glucose 1-acetic acid ester is converted primarily into the alpha- and beta-anomers of 2-O-acetyl D-glucose | Leuconostoc mesenteroides | 2-O-acetyl D-glucose + ? | - |
? | |
| alpha-D-glucose 1-phosphate + arsenate | - |
Leuconostoc mesenteroides | ? | - |
? | |
| additional information | regio- and stereoselective formation of alpha-glucose 1-acetic acid ester by mutant E237Q, NMR product determination, overview | Leuconostoc mesenteroides | ? | - |
? | |
| sucrose + phosphate | - |
Leuconostoc mesenteroides | D-fructose + alpha-D-glucose 1-phosphate | - |
r | |
| sucrose + phosphate | enzyme deglucosylation to an anionic nucleophile takes place with Glu237 protonated or unprotonated. Enzymatically formed alpha-glucose 1-esters decompose spontaneously via acyl group migration and hydrolysis | Leuconostoc mesenteroides | D-fructose + alpha-D-glucose 1-phosphate | - |
r | |
Subunits
| Subunits | Comment | Organism |
|---|---|---|
| ? | x * 55700, SDS-PAGE | Leuconostoc mesenteroides |
Synonyms
| Synonyms | Comment | Organism |
|---|---|---|
| SPase | - |
Leuconostoc mesenteroides |
Temperature Optimum [°C]
| Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
|---|---|---|---|
| 30 | - |
assay at | Leuconostoc mesenteroides |
Turnover Number [1/s]
| Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|
| 0.00063 | - |
alpha-D-glucose 1-phosphate | pH 7.0, 30°C, synthesis, mutant E237Q | Leuconostoc mesenteroides | |
| 0.0016 | - |
sucrose | pH 7.0, 30°C, phosphorolysis, mutant E237Q | Leuconostoc mesenteroides | |
| 72 | - |
D-fructose | pH 7.0, 30°C, synthesis, wild-type enzyme | Leuconostoc mesenteroides | |
| 91 | - |
alpha-D-glucose 1-phosphate | pH 7.0, 30°C, synthesis, wild-type enzyme | Leuconostoc mesenteroides | |
| 165 | - |
sucrose | pH 7.0, 30°C, phosphorolysis, wild-type enzyme | Leuconostoc mesenteroides | |
| 195 | - |
phosphate | pH 7.0, 30°C, phosphorolysis, wild-type enzyme | Leuconostoc mesenteroides | |
pH Optimum
| pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
|---|---|---|---|
| additional information | - |
deglucosylation of mutant E237Q is pH-independent | Leuconostoc mesenteroides |
| 5.6 | 7.2 | optimal pH for glycosylation | Leuconostoc mesenteroides |
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