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Literature summary extracted from

  • Cho, C.B.; Park, D.Y.; Lee, S.B.
    Effect of C-terminal domain truncation of Thermus thermophilus trehalose synthase on its substrate specificity (2017), Enzyme Microb. Technol., 96, 121-126 .
    View publication on PubMed

Cloned(Commentary)

EC Number Cloned (Comment) Organism
5.4.99.16 recombinant expression of His6-tagged wild-type and mutant enzymes in Escherichia coli strain BL21(DE3) Thermus thermophilus

Protein Variants

EC Number Protein Variants Comment Organism
5.4.99.16 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]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
5.4.99.16 additional information
-
additional information Michaelis-Menten kinetics of recombinant His6-tagged wild-type and mutant enzymes Thermus thermophilus
5.4.99.16 48
-
maltose recombinant His6-tagged wild-type enzyme, pH 6.5, 50°C Thermus thermophilus
5.4.99.16 62
-
alpha,alpha-trehalose recombinant His6-tagged wild-type enzyme, pH 6.5, 50°C Thermus thermophilus
5.4.99.16 77
-
maltose recombinant His6-tagged mutant DM2, pH 6.5, 50°C Thermus thermophilus
5.4.99.16 128
-
maltose recombinant His6-tagged mutant DM1, pH 6.5, 50°C Thermus thermophilus
5.4.99.16 456
-
alpha,alpha-trehalose recombinant His6-tagged mutant DM2, pH 6.5, 50°C Thermus thermophilus
5.4.99.16 1551
-
alpha,alpha-trehalose recombinant His6-tagged mutant DM1, pH 6.5, 50°C Thermus thermophilus

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
5.4.99.16 maltose Thermus thermophilus
-
alpha,alpha-trehalose
-
r
5.4.99.16 maltose Thermus thermophilus ATCC 33923
-
alpha,alpha-trehalose
-
r

Organism

EC Number Organism UniProt Comment Textmining
5.4.99.16 Thermus thermophilus Q7WUI5
-
-
5.4.99.16 Thermus thermophilus ATCC 33923 Q7WUI5
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
5.4.99.16 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)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
5.4.99.16 alpha,alpha-trehalose
-
Thermus thermophilus maltose
-
r
5.4.99.16 alpha,alpha-trehalose
-
Thermus thermophilus ATCC 33923 maltose
-
r
5.4.99.16 maltose
-
Thermus thermophilus alpha,alpha-trehalose
-
r
5.4.99.16 maltose
-
Thermus thermophilus ATCC 33923 alpha,alpha-trehalose
-
r
5.4.99.16 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 ?
-
?
5.4.99.16 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

EC Number Synonyms Comment Organism
5.4.99.16 Trehalose synthase
-
Thermus thermophilus
5.4.99.16 TtTS
-
Thermus thermophilus

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
5.4.99.16 50
-
assay at Thermus thermophilus

Temperature Stability [°C]

EC Number Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
5.4.99.16 100
-
10 min, inactivation Thermus thermophilus

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
5.4.99.16 6.5
-
assay at Thermus thermophilus

General Information

EC Number General Information Comment Organism
5.4.99.16 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
5.4.99.16 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