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Literature summary for 2.1.1.365 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
Search for more literature for 2.1.1.365

Cloned(Commentary)

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

Crystallization (Commentary)

Crystallization (Comment) Organism
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

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

Inhibitors

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

KM Value [mM]

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

Metals/Ions

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

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
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
-
 ?
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

Organism UniProt Comment Textmining
Mycolicibacterium hassiacum K5B7F3
-
-
Mycolicibacterium hassiacum DSM 44199 K5B7F3
-
-

Purification (Commentary)

Purification (Comment) Organism
HisTrap column chromatography Mycolicibacterium hassiacum

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
additional information the enzyme is inactive against mannose, the unmethylated dimannoside alpha-1,4-mannobiose, or 3-O-methylmannose Mycolicibacterium hassiacum ?
-
-
additional information the enzyme is inactive against mannose, the unmethylated dimannoside alpha-1,4-mannobiose, or 3-O-methylmannose Mycolicibacterium hassiacum DSM 44199 ?
-
-
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
-
 ?
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
-
 ?
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
-
 ?
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

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

Temperature Optimum [°C]

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

Turnover Number [1/s]

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

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
7.5
-
-
 Mycolicibacterium hassiacum

pH Range

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

kcat/KM [mM/s]

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