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Literature summary for 5.4.99.11 extracted from
- Mechanism-oriented redesign of an isomaltulose synthase to an isomelezitose synthase by site-directed mutagenesis (2012), ChemBioChem, 13, 149-156.
Application
| Application | Comment | Organism |
|---|---|---|
| synthesis | immobilized cells of Protaminobacter rubrum strain CBS 574.77 are commonly used for the synthesis of isomaltulose from sucrose on an industrial scale | Serratia plymuthica |
Cloned(Commentary)
| Cloned (Comment) | Organism |
|---|---|
| gene smuA, sequence comparison, expression of wild-type and mutant enzymes | Serratia plymuthica |
Protein Variants
| Protein Variants | Comment | Organism |
|---|---|---|
| E428D | site-directed mutagenesis, the mutant shows highly reduced activity compared to the wild-type enzyme | Serratia plymuthica |
| F297A | site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme | Serratia plymuthica |
| F297P | site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme | Serratia plymuthica |
| F297P/R333K | site-directed mutagenesis, the mutant shows highly reduced activity compared to the wild-type enzyme | Serratia plymuthica |
| F321A | site-directed mutagenesis, the mutant shows only hydrolytic activity | Serratia plymuthica |
| F321A/F319A | site-directed mutagenesis, the mutant shows highly reduced activity compared to the wild-type enzyme | Serratia plymuthica |
| additional information | engineering of the enzyme to exhibit isomelezitose synthase activity for production of the probiotically nutrial component, overview | Serratia plymuthica |
| R333K | site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme | Serratia plymuthica |
| R456K | site-directed mutagenesis, inactive mutant | Serratia plymuthica |
KM Value [mM]
| KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|
| additional information | - |
additional information | Michaelis-Menten kinetics of wild-type and mutant enzymes, overview | Serratia plymuthica | |
| 0.022 | - |
sucrose | pH 7.0, 30°C, recombinant wild-type enzyme | Serratia plymuthica |  |
Natural Substrates/ Products (Substrates)
| Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| additional information | Serratia plymuthica | the enzyme catalyzes the isomerization of sucrose (alpha-D-glucosylpyranosyl-1,2-beta-D-fructofuranose) to trehalulose (alpha-D-glucosylpyranosyl-1,1-D-fructofuranose) and isomaltulose (alpha-D-glucosylpyranosyl-1,6-D-fructofuranose) | ? | - |
? | |
| additional information | Serratia plymuthica CBS 574.77 | the enzyme catalyzes the isomerization of sucrose (alpha-D-glucosylpyranosyl-1,2-beta-D-fructofuranose) to trehalulose (alpha-D-glucosylpyranosyl-1,1-D-fructofuranose) and isomaltulose (alpha-D-glucosylpyranosyl-1,6-D-fructofuranose) | ? | - |
? | |
| sucrose | Serratia plymuthica | - |
6-O-alpha-D-Glucopyranosyl-D-fructofuranose | - |
? | |
| sucrose | Serratia plymuthica | - |
1-O-alpha-D-glucopyranosyl-D-fructofuranose + alpha-D-glucosylpyranosyl-1,6-D-fructofuranose | - |
? | |
| sucrose | Serratia plymuthica CBS 574.77 | - |
6-O-alpha-D-Glucopyranosyl-D-fructofuranose | - |
? | |
| sucrose | Serratia plymuthica CBS 574.77 | - |
1-O-alpha-D-glucopyranosyl-D-fructofuranose + alpha-D-glucosylpyranosyl-1,6-D-fructofuranose | - |
? | |
Organism
| Organism | UniProt | Comment | Textmining |
|---|---|---|---|
| Serratia plymuthica | D0VX20 | gene smuA | - |
| Serratia plymuthica CBS 574.77 | D0VX20 | gene smuA | - |
Specific Activity [micromol/min/mg]
| Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
|---|---|---|---|
| 0.0072 | - |
pH 7.0, 30°C, purified recombinant mutant F297A+P | Serratia plymuthica |
| 0.046 | - |
pH 7.0, 30°C, purified recombinant mutant F297A | Serratia plymuthica |
| 0.14 | - |
pH 7.0, 30°C, purified recombinant mutant R333K | Serratia plymuthica |
| 1.1 | - |
pH 7.0, 30°C, purified recombinant wild-type enzyme | Serratia plymuthica |
Substrates and Products (Substrate)
| Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| additional information | the enzyme catalyzes the isomerization of sucrose (alpha-D-glucosylpyranosyl-1,2-beta-D-fructofuranose) to trehalulose (alpha-D-glucosylpyranosyl-1,1-D-fructofuranose) and isomaltulose (alpha-D-glucosylpyranosyl-1,6-D-fructofuranose) | Serratia plymuthica | ? | - |
? | |
| additional information | the enzyme catalyzes the isomerization of sucrose (alpha-D-glucosylpyranosyl-1,2-beta-D-fructofuranose) to trehalulose (alpha-D-glucosylpyranosyl-1,1-D-fructofuranose) and isomaltulose (alpha-D-glucosylpyranosyl-1,6-D-fructofuranose) | Serratia plymuthica CBS 574.77 | ? | - |
? | |
| sucrose | - |
Serratia plymuthica | 6-O-alpha-D-Glucopyranosyl-D-fructofuranose | - |
? | |
| sucrose | isomerization of sucrose to isomaltulose is a highly efficient reaction, the formation of the beta-glucopyranosyl-enzyme complex followed by the nucleophilic attack of the 6'-OH group of fructose is concerted | Serratia plymuthica | 6-O-alpha-D-Glucopyranosyl-D-fructofuranose | - |
? | |
| sucrose | - |
Serratia plymuthica CBS 574.77 | 6-O-alpha-D-Glucopyranosyl-D-fructofuranose | - |
? | |
| sucrose | isomerization of sucrose to isomaltulose is a highly efficient reaction, the formation of the beta-glucopyranosyl-enzyme complex followed by the nucleophilic attack of the 6'-OH group of fructose is concerted | Serratia plymuthica CBS 574.77 | 6-O-alpha-D-Glucopyranosyl-D-fructofuranose | - |
? | |
| sucrose | - |
Serratia plymuthica | 1-O-alpha-D-glucopyranosyl-D-fructofuranose + alpha-D-glucosylpyranosyl-1,6-D-fructofuranose | - |
? | |
| sucrose | activity involves residues D241, E295, H368, and D369 | Serratia plymuthica | 1-O-alpha-D-glucopyranosyl-D-fructofuranose + alpha-D-glucosylpyranosyl-1,6-D-fructofuranose | - |
? | |
| sucrose | - |
Serratia plymuthica CBS 574.77 | 1-O-alpha-D-glucopyranosyl-D-fructofuranose + alpha-D-glucosylpyranosyl-1,6-D-fructofuranose | - |
? | |
| sucrose | activity involves residues D241, E295, H368, and D369 | Serratia plymuthica CBS 574.77 | 1-O-alpha-D-glucopyranosyl-D-fructofuranose + alpha-D-glucosylpyranosyl-1,6-D-fructofuranose | - |
? | |
Synonyms
| Synonyms | Comment | Organism |
|---|---|---|
| SmuA | - |
Serratia plymuthica |
| sucrose isomerase | - |
Serratia plymuthica |
Temperature Optimum [°C]
| Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
|---|---|---|---|
| 30 | - |
assay at | Serratia plymuthica |
Turnover Number [1/s]
| Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|
| 0.2 | - |
sucrose | pH 7.0, 30°C, purified recombinant mutant F297A+P | Serratia plymuthica | |
| 1.3 | - |
sucrose | pH 7.0, 30°C, purified recombinant mutant F297A | Serratia plymuthica | |
| 3.8 | - |
sucrose | pH 7.0, 30°C, purified recombinant mutant R333K | Serratia plymuthica | |
| 30 | - |
sucrose | pH 7.0, 30°C, recombinant wild-type enzyme | Serratia plymuthica | |
pH Optimum
| pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
|---|---|---|---|
| 7 | - |
assay at | Serratia plymuthica |
General Information
| General Information | Comment | Organism |
|---|---|---|
| additional information | enzyme-substrate complex, docking study with sucrose and fructose | Serratia plymuthica |
kcat/KM [mM/s]
| kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|
| 1.4 | - |
sucrose | pH 7.0, 30°C, recombinant wild-type enzyme | Serratia plymuthica | |
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