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Literature summary for 4.2.1.154 extracted from

  • Thompson, T.B.; Katayama, K.; Watanabe, K.; Hutchinson, C.R.; Rayment, I.
    Structural and functional analysis of tetracenomycin F2 cyclase from Streptomyces glaucescens. A type II polyketide cyclase (2004), J. Biol. Chem., 279, 37956-37963.
    View publication on PubMed
Search for more literature for 4.2.1.154

Cloned(Commentary)

Cloned (Comment) Organism
expressed in Escherichia coli BL21(DE3) cells Streptomyces glaucescens

Crystallization (Commentary)

Crystallization (Comment) Organism
macroseeding method, using 1.4 M (NH4)2SO4, 50 mM HEPES, pH 7.5 Streptomyces glaucescens

Protein Variants

Protein Variants Comment Organism
D27N the mutant shows 14% activity compared to the wild type enzyme Streptomyces glaucescens
H26A the mutant shows 15% activity compared to the wild type enzyme Streptomyces glaucescens
H26Q the mutant shows 91% activity compared to the wild type enzyme Streptomyces glaucescens
H51A the mutant shows 16% activity compared to the wild type enzyme Streptomyces glaucescens
H51Q the mutant shows 96% activity compared to the wild type enzyme Streptomyces glaucescens
R40G the mutant shows 10% activity compared to the wild type enzyme Streptomyces glaucescens
R40K the mutant shows 16% activity compared to the wild type enzyme Streptomyces glaucescens

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.155
-
 tetracenomycin F2 wild type enzyme, at 30°C in 0.1 M Tris-HCl, pH 7.5 Streptomyces glaucescens
0.191
-
 tetracenomycin F2 mutant enzyme H51Q, at 30°C in 0.1 M Tris-HCl, pH 7.5 Streptomyces glaucescens
0.203
-
 tetracenomycin F2 mutant enzyme H26Q, at 30°C in 0.1 M Tris-HCl, pH 7.5 Streptomyces glaucescens
0.732
-
 tetracenomycin F2 mutant enzyme H26A, at 30°C in 0.1 M Tris-HCl, pH 7.5 Streptomyces glaucescens
0.816
-
 tetracenomycin F2 mutant enzyme R40K, at 30°C in 0.1 M Tris-HCl, pH 7.5 Streptomyces glaucescens
0.892
-
 tetracenomycin F2 mutant enzyme H51A, at 30°C in 0.1 M Tris-HCl, pH 7.5 Streptomyces glaucescens
1.05
-
 tetracenomycin F2 mutant enzymeD27N, at 30°C in 0.1 M Tris-HCl, pH 7.5 Streptomyces glaucescens
1.2
-
 tetracenomycin F2 mutant enzyme R40G, at 30°C in 0.1 M Tris-HCl, pH 7.5 Streptomyces glaucescens

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
tetracenomycin F2 Streptomyces glaucescens
-
 tetracenomycin F1 + H2O
-
 ?

Organism

Organism UniProt Comment Textmining
Streptomyces glaucescens P39890
-
-

Purification (Commentary)

Purification (Comment) Organism
Q-Sepharose column chromatography, phenyl-Sepharose column chromatography, and MonoQ column chromatography Streptomyces glaucescens

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
tetracenomycin F2
-
 Streptomyces glaucescens tetracenomycin F1 + H2O
-
 ?

Subunits

Subunits Comment Organism
dimer
-
 Streptomyces glaucescens

Synonyms

Synonyms Comment Organism
tcmI
-
 Streptomyces glaucescens

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
0.0146
-
 tetracenomycin F2 mutant enzyme R40G, at 30°C in 0.1 M Tris-HCl, pH 7.5 Streptomyces glaucescens
0.0192
-
 tetracenomycin F2 mutant enzyme H26A, at 30°C in 0.1 M Tris-HCl, pH 7.5 Streptomyces glaucescens
0.0198
-
 tetracenomycin F2 mutant enzymeD27N, at 30°C in 0.1 M Tris-HCl, pH 7.5 Streptomyces glaucescens
0.0219
-
 tetracenomycin F2 mutant enzyme H51A, at 30°C in 0.1 M Tris-HCl, pH 7.5 Streptomyces glaucescens
0.025
-
 tetracenomycin F2 mutant enzyme R40K, at 30°C in 0.1 M Tris-HCl, pH 7.5 Streptomyces glaucescens
0.131
-
 tetracenomycin F2 mutant enzyme H26Q, at 30°C in 0.1 M Tris-HCl, pH 7.5 Streptomyces glaucescens
0.136
-
 tetracenomycin F2 mutant enzyme H51Q, at 30°C in 0.1 M Tris-HCl, pH 7.5 Streptomyces glaucescens
0.142
-
 tetracenomycin F2 wild type enzyme, at 30°C in 0.1 M Tris-HCl, pH 7.5 Streptomyces glaucescens

kcat/KM [mM/s]

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
0.0121
-
 tetracenomycin F2 mutant enzyme R40G, at 30°C in 0.1 M Tris-HCl, pH 7.5 Streptomyces glaucescens
0.0188
-
 tetracenomycin F2 mutant enzymeD27N, at 30°C in 0.1 M Tris-HCl, pH 7.5 Streptomyces glaucescens
0.0245
-
 tetracenomycin F2 mutant enzyme H51A, at 30°C in 0.1 M Tris-HCl, pH 7.5 Streptomyces glaucescens
0.0262
-
 tetracenomycin F2 mutant enzyme H26A, at 30°C in 0.1 M Tris-HCl, pH 7.5 Streptomyces glaucescens
0.0306
-
 tetracenomycin F2 mutant enzyme R40K, at 30°C in 0.1 M Tris-HCl, pH 7.5 Streptomyces glaucescens
0.645
-
 tetracenomycin F2 mutant enzyme H26Q, at 30°C in 0.1 M Tris-HCl, pH 7.5 Streptomyces glaucescens
0.712
-
 tetracenomycin F2 mutant enzyme H51Q, at 30°C in 0.1 M Tris-HCl, pH 7.5 Streptomyces glaucescens
0.916
-
 tetracenomycin F2 wild type enzyme, at 30°C in 0.1 M Tris-HCl, pH 7.5 Streptomyces glaucescens