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Literature summary extracted from

  • Li, C.H.; Tu, S.C.
    Active site hydrophobicity is critical to the bioluminescence activity of Vibrio harveyi luciferase (2005), Biochemistry, 44, 12970-12977.
    View publication on PubMed

Cloned(Commentary)

EC Number Cloned (Comment) Organism
1.14.14.3 overexpression of wild-type and mutant enzymes in Escherichia coli strain JM101 Vibrio harveyi

Protein Variants

EC Number Protein Variants Comment Organism
1.14.14.3 alphaF114A site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme due to reduced hydrophobicity of the active site Vibrio harveyi
1.14.14.3 alphaF114D site-directed mutagenesis, the mutant shows highly reduced activity and increased Km compared to the wild-type enzyme due to reduced hydrophobicity of the active site Vibrio harveyi
1.14.14.3 alphaF114S site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme due to reduced hydrophobicity of the active site Vibrio harveyi
1.14.14.3 alphaF114Y site-directed mutagenesis, the mutant shows slightly reduced activity compared to the wild-type enzyme Vibrio harveyi
1.14.14.3 alphaF117A site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme due to reduced hydrophobicity of the active site Vibrio harveyi
1.14.14.3 alphaF117D site-directed mutagenesis, the mutant shows highly reduced activity and increased Km compared to the wild-type enzyme due to reduced hydrophobicity of the active site Vibrio harveyi
1.14.14.3 alphaF117S site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme due to reduced hydrophobicity of the active site Vibrio harveyi
1.14.14.3 alphaF117Y site-directed mutagenesis, the mutant shows slightly reduced activity compared to the wild-type enzyme Vibrio harveyi
1.14.14.3 alphaF327A site-directed mutagenesis, mutant activity is similar to the wild-type enzyme Vibrio harveyi
1.14.14.3 alphaF46A site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme due to reduced hydrophobicity of the active site Vibrio harveyi
1.14.14.3 alphaF46D site-directed mutagenesis, the mutant shows highly reduced activity and increased Km compared to the wild-type enzyme due to reduced hydrophobicity of the active site Vibrio harveyi
1.14.14.3 alphaF46S site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme due to reduced hydrophobicity of the active site Vibrio harveyi
1.14.14.3 alphaF46Y site-directed mutagenesis, the mutant shows slightly reduced activity compared to the wild-type enzyme Vibrio harveyi
1.14.14.3 alphaF49A site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme due to reduced hydrophobicity of the active site Vibrio harveyi
1.14.14.3 alphaF49D site-directed mutagenesis, the mutant shows highly reduced activity and increased Km compared to the wild-type enzyme due to reduced hydrophobicity of the active site Vibrio harveyi
1.14.14.3 alphaF49S site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme due to reduced hydrophobicity of the active site Vibrio harveyi
1.14.14.3 alphaF49Y site-directed mutagenesis, the mutant shows slightly reduced activity compared to the wild-type enzyme Vibrio harveyi
1.14.14.3 alphaF6A site-directed mutagenesis, mutant activity is similar to the wild-type enzyme Vibrio harveyi

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
1.14.14.3 additional information
-
 additional information stopped-flow kinetics of wild-type and mutant enzymes Vibrio harveyi
1.14.14.3 0.0002
-
 FMNH2 pH 7.0, 23°C, recombinant mutant F114S Vibrio harveyi
1.14.14.3 0.0003
-
 FMNH2 pH 7.0, 23°C, recombinant mutants F46Y, F114A, F117Y, and F114Y Vibrio harveyi
1.14.14.3 0.0006
-
 FMNH2 pH 7.0, 23°C, recombinant mutant F49Y Vibrio harveyi
1.14.14.3 0.0006
-
 FMNH2 pH 7.0, 23°C, recombinant wild-type enzyme Vibrio harveyi
1.14.14.3 0.0007
-
 FMNH2 pH 7.0, 23°C, recombinant mutant F49S Vibrio harveyi
1.14.14.3 0.001
-
 FMNH2 pH 7.0, 23°C, recombinant mutant F114D Vibrio harveyi
1.14.14.3 0.00101
-
 decanal pH 7.0, 23°C, recombinant mutant F49Y Vibrio harveyi
1.14.14.3 0.0013
-
 FMNH2 pH 7.0, 23°C, recombinant mutant F117S Vibrio harveyi
1.14.14.3 0.0013
-
 decanal pH 7.0, 23°C, recombinant mutant F46A Vibrio harveyi
1.14.14.3 0.0013
-
 FMNH2 pH 7.0, 23°C, recombinant mutant F49D Vibrio harveyi
1.14.14.3 0.0015
-
 FMNH2 pH 7.0, 23°C, recombinant mutant F117A Vibrio harveyi
1.14.14.3 0.0016
-
 decanal pH 7.0, 23°C, recombinant wild-type enzyme Vibrio harveyi
1.14.14.3 0.0024
-
 decanal pH 7.0, 23°C, recombinant mutant F117S Vibrio harveyi
1.14.14.3 0.0024
-
 decanal pH 7.0, 23°C, recombinant mutant F46Y Vibrio harveyi
1.14.14.3 0.0026
-
 decanal pH 7.0, 23°C, recombinant mutant F46S Vibrio harveyi
1.14.14.3 0.0027
-
 decanal pH 7.0, 23°C, recombinant mutant F114Y Vibrio harveyi
1.14.14.3 0.0027
-
 FMNH2 pH 7.0, 23°C, recombinant mutant F49A Vibrio harveyi
1.14.14.3 0.003
-
 decanal pH 7.0, 23°C, recombinant mutants F49D, F117A, and F49A Vibrio harveyi
1.14.14.3 0.0031
-
 decanal pH 7.0, 23°C, recombinant mutant F117D Vibrio harveyi
1.14.14.3 0.0032
-
 decanal pH 7.0, 23°C, recombinant mutant F46D Vibrio harveyi
1.14.14.3 0.0033
-
 decanal pH 7.0, 23°C, recombinant mutant F49S Vibrio harveyi
1.14.14.3 0.0043
-
 FMNH2 pH 7.0, 23°C, recombinant mutant F46S Vibrio harveyi
1.14.14.3 0.0051
-
 decanal pH 7.0, 23°C, recombinant mutant F117Y Vibrio harveyi
1.14.14.3 0.0073
-
 decanal pH 7.0, 23°C, recombinant mutant F114D Vibrio harveyi
1.14.14.3 0.0083
-
 decanal pH 7.0, 23°C, recombinant mutant F114S Vibrio harveyi
1.14.14.3 0.0093
-
 decanal pH 7.0, 23°C, recombinant mutant F114A Vibrio harveyi
1.14.14.3 0.0123
-
 FMNH2 pH 7.0, 23°C, recombinant mutant F46A Vibrio harveyi
1.14.14.3 0.0148
-
 FMNH2 pH 7.0, 23°C, recombinant mutant F46D Vibrio harveyi
1.14.14.3 0.052
-
 FMNH2 pH 7.0, 23°C, recombinant mutant F117D Vibrio harveyi

Organism

EC Number Organism UniProt Comment Textmining
1.14.14.3 Vibrio harveyi
-
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
1.14.14.3 recombinant wild-type and mutant enzymes from Escherichia coli strain JM101 to homogeneity Vibrio harveyi

Reaction

EC Number Reaction Comment Organism Reaction ID
1.14.14.3 a long-chain aldehyde + FMNH2 + O2 = a long-chain fatty acid + FMN + H2O + hnu reaction mechanism via 4a-hydroperoxyflavin intermediates II-IV and H2O2 release, overview, catalytically important active site residues of the alpha-subunit are alphaF46, alphaF49, alphaF114, and alphaF117, alphaF46 and alphaF117 also appear to be involved in the binding of reduced flavin substrate Vibrio harveyi
a

Specific Activity [micromol/min/mg]

EC Number Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
1.14.14.3 additional information
-
 quantum yield of wild-type and mutant enzymes, overview Vibrio harveyi

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
1.14.14.3 decanal + FMNH2 + O2
-
 Vibrio harveyi decanoate + FMN + H2O + hv
-
 ir
1.14.14.3 additional information active site hydrophobicity is critical to the bioluminescence activity of Vibrio harveyi luciferase Vibrio harveyi ?
-
 ?

Subunits

EC Number Subunits Comment Organism
1.14.14.3 dimer alphabeta heterodimer containing a single active site at a cleft in the alpha subunit Vibrio harveyi

Synonyms

EC Number Synonyms Comment Organism
1.14.14.3 luciferase
-
 Vibrio harveyi
1.14.14.3 Vibrio harveyi luciferase
-
 Vibrio harveyi

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
1.14.14.3 23
-
 assay at Vibrio harveyi

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
1.14.14.3 7
-
 assay at Vibrio harveyi

Cofactor

EC Number Cofactor Comment Organism Structure
1.14.14.3 FMNH2
-
 Vibrio harveyi