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Literature summary 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

Cloned(Commentary)

EC Number Cloned (Comment) Organism
2.1.1.303 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)

EC Number Crystallization (Comment) Organism
2.1.1.303 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

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

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
2.1.1.303 0.206
-
 2,7-dihydroxy-5-methyl-1-naphthoate pH 7.5, 25°C, wild-type enzyme Streptomyces carzinostaticus
2.1.1.303 0.319
-
 2,7-dihydroxy-5-methyl-1-naphthoate pH 7.5, 25°C, mutant enzyme R11K Streptomyces carzinostaticus
2.1.1.303 0.4
-
 2,7-dihydroxy-5-methyl-1-naphthoate pH 7.5, 25°C, mutant enzyme R11W Streptomyces carzinostaticus
2.1.1.303 0.419
-
 2,7-dihydroxy-5-methyl-1-naphthoate pH 7.5, 25°C, mutant enzyme R11A Streptomyces carzinostaticus
2.1.1.303 0.649
-
 2,7-dihydroxy-5-methyl-1-naphthoate pH 7.5, 25°C, mutant enzyme Y293I Streptomyces carzinostaticus

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
2.1.1.303 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

EC Number Organism UniProt Comment Textmining
2.1.1.303 Streptomyces carzinostaticus Q84HC8
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
2.1.1.303
-
 Streptomyces carzinostaticus

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
2.1.1.303 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
-
 ?
2.1.1.303 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

EC Number Synonyms Comment Organism
2.1.1.303 NcsB1
-
 Streptomyces carzinostaticus

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
2.1.1.303 25
-
 assay at Streptomyces carzinostaticus

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
2.1.1.303 0.01
-
 2,7-dihydroxy-5-methyl-1-naphthoate pH 7.5, 25°C, mutant enzyme R11W Streptomyces carzinostaticus
2.1.1.303 0.0115
-
 2,7-dihydroxy-5-methyl-1-naphthoate pH 7.5, 25°C, wild-type enzyme Streptomyces carzinostaticus
2.1.1.303 0.013
-
 2,7-dihydroxy-5-methyl-1-naphthoate pH 7.5, 25°C, mutant enzyme Y293I Streptomyces carzinostaticus
2.1.1.303 0.02
-
 2,7-dihydroxy-5-methyl-1-naphthoate pH 7.5, 25°C, mutant enzyme R11A Streptomyces carzinostaticus
2.1.1.303 0.02
-
 2,7-dihydroxy-5-methyl-1-naphthoate pH 7.5, 25°C, mutant enzyme R11K Streptomyces carzinostaticus

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
2.1.1.303 7.5
-
 assay at Streptomyces carzinostaticus

General Information

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

kcat/KM [mM/s]

EC Number kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
2.1.1.303 0.02
-
 2,7-dihydroxy-5-methyl-1-naphthoate pH 7.5, 25°C, mutant enzyme Y293I Streptomyces carzinostaticus
2.1.1.303 0.025
-
 2,7-dihydroxy-5-methyl-1-naphthoate pH 7.5, 25°C, mutant enzyme R11W Streptomyces carzinostaticus
2.1.1.303 0.048
-
 2,7-dihydroxy-5-methyl-1-naphthoate pH 7.5, 25°C, mutant enzyme R11A Streptomyces carzinostaticus
2.1.1.303 0.06
-
 2,7-dihydroxy-5-methyl-1-naphthoate pH 7.5, 25°C, wild-type enzyme Streptomyces carzinostaticus
2.1.1.303 0.063
-
 2,7-dihydroxy-5-methyl-1-naphthoate pH 7.5, 25°C, mutant enzyme TR11K Streptomyces carzinostaticus