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Literature summary for 2.1.1.303 extracted from

  • Cooke, H.A.; Guenther, E.L.; Luo, Y.; Shen, B.; Bruner, S.D.
    Molecular basis of substrate promiscuity for the SAM-dependent O-methyltransferase NcsB1, involved in the biosynthesis of the enediyne antitumor antibiotic neocarzinostatin (2009), Biochemistry, 48, 9590-9598.
    View publication on PubMedView publication on EuropePMC
Search for more literature for 2.1.1.303

Cloned(Commentary)

Cloned (Comment) Organism
the enzyme is overproduced as an N-terminal His6-tagged fusion protein using expression plasmid pBS5039 in Escherichia coli BL21(DE3) Streptomyces carzinostaticus

Crystallization (Commentary)

Crystallization (Comment) Organism
the enzyme is crystallized with a substituted naphthoic acid and/or S-adenosyl-L-methionine/S-adenosyl-L-homocysteine by the hanging drop vapor diffusion method Streptomyces carzinostaticus

Protein Variants

Protein Variants Comment Organism
R11A kcat/Km is 88% of the wild-type value Streptomyces carzinostaticus
R11K kcat/Km is 110% of the wild-type value Streptomyces carzinostaticus
R11W kcat/Km is 45% of the wild-type value Streptomyces carzinostaticus
Y293I kcat/Km is 35% of the wild-type value Streptomyces carzinostaticus

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.206
-
 2,7-dihydroxy-5-methyl-1-naphthoate pH 7.5, 25°C, wild-type enzyme Streptomyces carzinostaticus
0.319
-
 2,7-dihydroxy-5-methyl-1-naphthoate pH 7.5, 25°C, mutant enzyme R11K Streptomyces carzinostaticus
0.4
-
 2,7-dihydroxy-5-methyl-1-naphthoate pH 7.5, 25°C, mutant enzyme R11W Streptomyces carzinostaticus
0.419
-
 2,7-dihydroxy-5-methyl-1-naphthoate pH 7.5, 25°C, mutant enzyme R11A Streptomyces carzinostaticus
0.649
-
 2,7-dihydroxy-5-methyl-1-naphthoate pH 7.5, 25°C, mutant enzyme Y293I Streptomyces carzinostaticus

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
S-adenosyl-L-methionine + 2,7-dihydroxy-5-methyl-1-naphthoate Streptomyces carzinostaticus the enzyme is involved in the biosynthesis of the enediyne antitumor antibiotic neocarzinostatin S-adenosyl-L-homocysteine + 2-hydroxy-7-methoxy-5-methyl-1-naphthoate
-
 ?

Organism

Organism UniProt Comment Textmining
Streptomyces carzinostaticus Q84HC8
-
-

Purification (Commentary)

Purification (Comment) Organism
-
 Streptomyces carzinostaticus

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
S-adenosyl-L-methionine + 2,7-dihydroxy-5-methyl-1-naphthoate
-
 Streptomyces carzinostaticus S-adenosyl-L-homocysteine + 2-hydroxy-7-methoxy-5-methyl-1-naphthoate
-
 ?
S-adenosyl-L-methionine + 2,7-dihydroxy-5-methyl-1-naphthoate the enzyme is involved in the biosynthesis of the enediyne antitumor antibiotic neocarzinostatin Streptomyces carzinostaticus S-adenosyl-L-homocysteine + 2-hydroxy-7-methoxy-5-methyl-1-naphthoate
-
 ?

Synonyms

Synonyms Comment Organism
NcsB1
-
 Streptomyces carzinostaticus

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
25
-
 assay at Streptomyces carzinostaticus

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
0.01
-
 2,7-dihydroxy-5-methyl-1-naphthoate pH 7.5, 25°C, mutant enzyme R11W Streptomyces carzinostaticus
0.0115
-
 2,7-dihydroxy-5-methyl-1-naphthoate pH 7.5, 25°C, wild-type enzyme Streptomyces carzinostaticus
0.013
-
 2,7-dihydroxy-5-methyl-1-naphthoate pH 7.5, 25°C, mutant enzyme Y293I Streptomyces carzinostaticus
0.02
-
 2,7-dihydroxy-5-methyl-1-naphthoate pH 7.5, 25°C, mutant enzyme R11A Streptomyces carzinostaticus
0.02
-
 2,7-dihydroxy-5-methyl-1-naphthoate pH 7.5, 25°C, mutant enzyme R11K Streptomyces carzinostaticus

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
7.5
-
 assay at Streptomyces carzinostaticus

General Information

General Information Comment Organism
physiological function the enzyme is involved in the biosynthesis of the enediyne antitumor antibiotic neocarzinostatin Streptomyces carzinostaticus

kcat/KM [mM/s]

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
0.02
-
 2,7-dihydroxy-5-methyl-1-naphthoate pH 7.5, 25°C, mutant enzyme Y293I Streptomyces carzinostaticus
0.025
-
 2,7-dihydroxy-5-methyl-1-naphthoate pH 7.5, 25°C, mutant enzyme R11W Streptomyces carzinostaticus
0.048
-
 2,7-dihydroxy-5-methyl-1-naphthoate pH 7.5, 25°C, mutant enzyme R11A Streptomyces carzinostaticus
0.06
-
 2,7-dihydroxy-5-methyl-1-naphthoate pH 7.5, 25°C, wild-type enzyme Streptomyces carzinostaticus
0.063
-
 2,7-dihydroxy-5-methyl-1-naphthoate pH 7.5, 25°C, mutant enzyme TR11K Streptomyces carzinostaticus