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Literature summary for 5.4.99.16 extracted from
- Effect of C-terminal domain truncation of Thermus thermophilus trehalose synthase on its substrate specificity (2017), Enzyme Microb. Technol., 96, 121-126 .
Cloned(Commentary)
| Cloned (Comment) | Organism |
|---|---|
| recombinant expression of His6-tagged wild-type and mutant enzymes in Escherichia coli strain BL21(DE3) | Thermus thermophilus |
Protein Variants
| Protein Variants | Comment | Organism |
|---|---|---|
| additional information | construction of two truncated enzymes (DM1 and DM2), which show lower maltose- and trehalose-converting activities and a different transglycosylation reaction mechanism compared to the wild-type enzyme. In the mutants, the glucose moiety cleaved from the maltose substrate is released from the enzyme and intercepted by external glucose oxidase, preventing the production of trehalose | Thermus thermophilus |
KM Value [mM]
| KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|
| additional information | - |
additional information | Michaelis-Menten kinetics of recombinant His6-tagged wild-type and mutant enzymes | Thermus thermophilus | |
| 48 | - |
maltose | recombinant His6-tagged wild-type enzyme, pH 6.5, 50°C | Thermus thermophilus |  |
| 62 | - |
alpha,alpha-trehalose | recombinant His6-tagged wild-type enzyme, pH 6.5, 50°C | Thermus thermophilus | |
| 77 | - |
maltose | recombinant His6-tagged mutant DM2, pH 6.5, 50°C | Thermus thermophilus | |
| 128 | - |
maltose | recombinant His6-tagged mutant DM1, pH 6.5, 50°C | Thermus thermophilus | |
| 456 | - |
alpha,alpha-trehalose | recombinant His6-tagged mutant DM2, pH 6.5, 50°C | Thermus thermophilus | |
| 1551 | - |
alpha,alpha-trehalose | recombinant His6-tagged mutant DM1, pH 6.5, 50°C | Thermus thermophilus | |
Natural Substrates/ Products (Substrates)
| Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| maltose | Thermus thermophilus | - |
alpha,alpha-trehalose | - |
r | |
| maltose | Thermus thermophilus ATCC 33923 | - |
alpha,alpha-trehalose | - |
r | |
Organism
| Organism | UniProt | Comment | Textmining |
|---|---|---|---|
| Thermus thermophilus | Q7WUI5 | - |
- |
| Thermus thermophilus ATCC 33923 | Q7WUI5 | - |
- |
Purification (Commentary)
| Purification (Comment) | Organism |
|---|---|
| recombinant His6-tagged wild-type and mutant enzymes from Escherichia coli strain BL21(DE3) by nickel affinity chromatography and ultrafiltration | Thermus thermophilus |
Substrates and Products (Substrate)
| Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| alpha,alpha-trehalose | - |
Thermus thermophilus | maltose | - |
r | |
| alpha,alpha-trehalose | - |
Thermus thermophilus ATCC 33923 | maltose | - |
r | |
| maltose | - |
Thermus thermophilus | alpha,alpha-trehalose | - |
r | |
| maltose | - |
Thermus thermophilus ATCC 33923 | alpha,alpha-trehalose | - |
r | |
| additional information | the wild-type enzyme retains the glucose moiety in the active site during the reaction to effectively produce trehalose. 13C NMR analysis is performed to identify the glycosidic structure of the purified transfer disaccharide product, where the carbon signals are compared with that of alpha-mannose. Activity analysis by thin-layer chromatography | Thermus thermophilus | ? | - |
? | |
| additional information | the wild-type enzyme retains the glucose moiety in the active site during the reaction to effectively produce trehalose. 13C NMR analysis is performed to identify the glycosidic structure of the purified transfer disaccharide product, where the carbon signals are compared with that of alpha-mannose. Activity analysis by thin-layer chromatography | Thermus thermophilus ATCC 33923 | ? | - |
? | |
Synonyms
| Synonyms | Comment | Organism |
|---|---|---|
| Trehalose synthase | - |
Thermus thermophilus |
| TtTS | - |
Thermus thermophilus |
Temperature Optimum [°C]
| Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
|---|---|---|---|
| 50 | - |
assay at | Thermus thermophilus |
Temperature Stability [°C]
| Temperature Stability Minimum [°C] | Temperature Stability Maximum [°C] | Comment | Organism |
|---|---|---|---|
| 100 | - |
10 min, inactivation | Thermus thermophilus |
pH Optimum
| pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
|---|---|---|---|
| 6.5 | - |
assay at | Thermus thermophilus |
General Information
| General Information | Comment | Organism |
|---|---|---|
| malfunction | the C-terminal domain of the three-domain-comprising trehalose synthase from Thermus thermophilus is truncated in order to study the effect on the enzyme's activity and substrate specificity. Two truncated enzymes (DM1 and DM2) show lower maltose- and trehalose-converting activities and a different transglycosylation reaction mechanism compared to the wild-type enzyme. In the mutants, the glucose moiety cleaved from the maltose substrate is released from the enzyme and intercepted by external glucose oxidase, preventing the production of trehalose. Mutant DM1 synthesizes much higher amounts of mannose-containing disaccharide trehalose analogue (Man-TA) than does the wild-type or mutant DM2. The mutant enzymes could be used to produce Man-TA, a postulatedinhibitor of gut disaccharidases | Thermus thermophilus |
| additional information | the C-terminal domain in the wild-type enzyme is important for retaining the glucose moiety of the substrate within the active site | Thermus thermophilus |
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