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Literature summary extracted from

  • Elamin, A.; Stehr, M.; Oehlmann, W.; Singh, M.
    The mycolyltransferase 85A, a putative drug target of Mycobacterium tuberculosis: Development of a novel assay and quantification of glycolipid-status of the mycobacterial cell wall (2009), J. Microbiol. Methods, 79, 358-363.
    View publication on PubMed

Application

EC Number Application Comment Organism
2.3.1.122 biotechnology an enzyme assay using the natural substrate trehalose dimycolate is developed. The colorimetric assay is based on the quantification of glucose from the degradation of trehalose, which is the product from catalytic activity of antigen 85A. This assay is suitable for robust high-throughput screening (HTS) for compound library screening against mycolyltransferase. The assay has a very low coefficient of variance (0.04) in 96-well plates and shows a Z' factor of 0.67-0.73, indicating the robustness of the assay Mycolicibacterium smegmatis

Cloned(Commentary)

EC Number Cloned (Comment) Organism
2.3.1.122 expressed in Escherichia coli as a His-tagged fusion protein Mycolicibacterium smegmatis

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
2.3.1.122 0.1296
-
 alpha,alpha-trehalose 6-monomycolate pH 5.7, 37°C Mycolicibacterium smegmatis

Localization

EC Number Localization Comment Organism GeneOntology No. Textmining

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
2.3.1.122 32000
-
 SDS-PAGE Mycolicibacterium smegmatis

Organism

EC Number Organism UniProt Comment Textmining
2.3.1.122 Mycolicibacterium smegmatis
-
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
2.3.1.122 using Ni-NTA chromatography and by applying a linear imidazole gradient from 10 to 500 mM. The final purification step contains gel filtration on a Superdex 200 column Mycolicibacterium smegmatis

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
2.3.1.122 alpha,alpha'-trehalose 6-monomycolate + alpha,alpha'-trehalose 6-monomycolate
-
 Mycolicibacterium smegmatis alpha,alpha'-trehalose 6,6'-dimycolate + alpha,alpha'-trehalose
-
 ?

Synonyms

EC Number Synonyms Comment Organism
2.3.1.122 antigen 85A
-
 Mycolicibacterium smegmatis
2.3.1.122 mycolyltransferase
-
 Mycolicibacterium smegmatis

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
2.3.1.122 37
-
 assay at Mycolicibacterium smegmatis

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
2.3.1.122 1.09
-
 alpha,alpha-trehalose 6-monomycolate pH 5.7, 37°C Mycolicibacterium smegmatis

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
2.3.1.122 5.7
-
 assay at Mycolicibacterium smegmatis