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Literature summary extracted from

  • Ripoll-Rozada, J.; Costa, M.; Manso, J.A.; Maranha, A.; Miranda, V.; Sequeira, A.; Ventura, M.R.; Macedo-Ribeiro, S.; Pereira, P.J.B.; Empadinhas, N.
    Biosynthesis of mycobacterial methylmannose polysaccharides requires a unique 1-O-methyltransferase specific for 3-O-methylated mannosides (2019), Proc. Natl. Acad. Sci. USA, 116, 835-844 .
    View publication on PubMedView publication on EuropePMC

Cloned(Commentary)

EC Number Cloned (Comment) Organism
2.1.1.365 expressed as His-tagged recombinant protein in Escherichia coli BL21(DE3) cells Mycolicibacterium hassiacum

Crystallization (Commentary)

EC Number Crystallization (Comment) Organism
2.1.1.365 in complex with S-adenosyl-L-homocysteine, sitting drop vapor diffusion method, using 0.1 M Tris-HCl pH 8.5, 0.2 M MgCl2, 20% (w/v) PEG 8000 or 0.1 M MES pH 6.5, and 1.6 M MgSO4 Mycolicibacterium hassiacum

Protein Variants

EC Number Protein Variants Comment Organism
2.1.1.365 E78A the mutant shows about 10% activity compared to the wild type enzyme Mycolicibacterium hassiacum
2.1.1.365 H144A inactive Mycolicibacterium hassiacum
2.1.1.365 H79A the mutant shows about 13% activity compared to the wild type enzyme Mycolicibacterium hassiacum

Inhibitors

EC Number Inhibitors Comment Organism Structure
2.1.1.365 EDTA the recombinant enzyme is inactivated by 10 mM EDTA Mycolicibacterium hassiacum

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
2.1.1.365 0.055
-
 S-adenosyl-L-methionine at pH 7.5 and 37°C Mycolicibacterium hassiacum
2.1.1.365 0.25
-
 3,3'-di-O-methyl-4alpha-mannobiose wild type enzyme, at pH 7.5 and 37°C Mycolicibacterium hassiacum
2.1.1.365 0.69
-
 3,3'-di-O-methyl-4alpha-mannobiose mutant enzyme E78A, at pH 7.5 and 37°C Mycolicibacterium hassiacum
2.1.1.365 0.78
-
 3,3'-di-O-methyl-4alpha-mannobiose mutant enzyme H79A, at pH 7.5 and 37°C Mycolicibacterium hassiacum

Metals/Ions

EC Number Metals/Ions Comment Organism Structure
2.1.1.365 Mg2+ required, 10 mM used in assay conditions Mycolicibacterium hassiacum

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
2.1.1.365 S-adenosyl-L-methionine + 3,3'-di-O-methyl-4alpha-mannobiose Mycolicibacterium hassiacum
-
 S-adenosyl-L-homocysteine + 1,3,3'-tri-O-methyl-4alpha-mannobiose
-
 ?
2.1.1.365 S-adenosyl-L-methionine + 3,3'-di-O-methyl-4alpha-mannobiose Mycolicibacterium hassiacum DSM 44199
-
 S-adenosyl-L-homocysteine + 1,3,3'-tri-O-methyl-4alpha-mannobiose
-
 ?

Organism

EC Number Organism UniProt Comment Textmining
2.1.1.365 Mycolicibacterium hassiacum K5B7F3
-
-
2.1.1.365 Mycolicibacterium hassiacum DSM 44199 K5B7F3
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
2.1.1.365 HisTrap column chromatography Mycolicibacterium hassiacum

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
2.1.1.365 additional information the enzyme is inactive against mannose, the unmethylated dimannoside alpha-1,4-mannobiose, or 3-O-methylmannose Mycolicibacterium hassiacum ?
-
-
2.1.1.365 additional information the enzyme is inactive against mannose, the unmethylated dimannoside alpha-1,4-mannobiose, or 3-O-methylmannose Mycolicibacterium hassiacum DSM 44199 ?
-
-
2.1.1.365 S-adenosyl-L-methionine + 3,3'-di-O-methyl-4alpha-mannobiose
-
 Mycolicibacterium hassiacum S-adenosyl-L-homocysteine + 1,3,3'-tri-O-methyl-4alpha-mannobiose
-
 ?
2.1.1.365 S-adenosyl-L-methionine + 3,3'-di-O-methyl-4alpha-mannobiose specific substrate Mycolicibacterium hassiacum S-adenosyl-L-homocysteine + 1,3,3'-tri-O-methyl-4alpha-mannobiose
-
 ?
2.1.1.365 S-adenosyl-L-methionine + 3,3'-di-O-methyl-4alpha-mannobiose
-
 Mycolicibacterium hassiacum DSM 44199 S-adenosyl-L-homocysteine + 1,3,3'-tri-O-methyl-4alpha-mannobiose
-
 ?
2.1.1.365 S-adenosyl-L-methionine + 3,3'-di-O-methyl-4alpha-mannobiose specific substrate Mycolicibacterium hassiacum DSM 44199 S-adenosyl-L-homocysteine + 1,3,3'-tri-O-methyl-4alpha-mannobiose
-
 ?

Synonyms

EC Number Synonyms Comment Organism
2.1.1.365 MET1
-
 Mycolicibacterium hassiacum
2.1.1.365 S-adenosyl-L-methionine-dependent sugar 1-O-methyltransferase
-
 Mycolicibacterium hassiacum

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
2.1.1.365 50 55
-
 Mycolicibacterium hassiacum

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
2.1.1.365 0.000033
-
 3,3'-di-O-methyl-4alpha-mannobiose mutant enzyme E78A, at pH 7.5 and 37°C Mycolicibacterium hassiacum
2.1.1.365 0.000033
-
 3,3'-di-O-methyl-4alpha-mannobiose mutant enzyme H79A, at pH 7.5 and 37°C Mycolicibacterium hassiacum
2.1.1.365 0.00062
-
 3,3'-di-O-methyl-4alpha-mannobiose wild type enzyme, at pH 7.5 and 37°C Mycolicibacterium hassiacum

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
2.1.1.365 7.5
-
-
 Mycolicibacterium hassiacum

pH Range

EC Number pH Minimum pH Maximum Comment Organism
2.1.1.365 6.5 9.5 the enzyme is active between pH 6.5 and 9.5 Mycolicibacterium hassiacum

kcat/KM [mM/s]

EC Number kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
2.1.1.365 0.00005
-
 3,3'-di-O-methyl-4alpha-mannobiose mutant enzyme E78A, at pH 7.5 and 37°C Mycolicibacterium hassiacum
2.1.1.365 0.00005
-
 3,3'-di-O-methyl-4alpha-mannobiose mutant enzyme H79A, at pH 7.5 and 37°C Mycolicibacterium hassiacum
2.1.1.365 0.0025
-
 3,3'-di-O-methyl-4alpha-mannobiose wild type enzyme, at pH 7.5 and 37°C Mycolicibacterium hassiacum