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Literature summary for 1.14.14.3 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
Search for more literature for 1.14.14.3

Cloned(Commentary)

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

Protein Variants

Protein Variants Comment Organism
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
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
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
alphaF114Y site-directed mutagenesis, the mutant shows slightly reduced activity compared to the wild-type enzyme Vibrio harveyi
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
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
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
alphaF117Y site-directed mutagenesis, the mutant shows slightly reduced activity compared to the wild-type enzyme Vibrio harveyi
alphaF327A site-directed mutagenesis, mutant activity is similar to the wild-type enzyme Vibrio harveyi
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
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
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
alphaF46Y site-directed mutagenesis, the mutant shows slightly reduced activity compared to the wild-type enzyme Vibrio harveyi
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
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
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
alphaF49Y site-directed mutagenesis, the mutant shows slightly reduced activity compared to the wild-type enzyme Vibrio harveyi
alphaF6A site-directed mutagenesis, mutant activity is similar to the wild-type enzyme Vibrio harveyi

KM Value [mM]

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

Organism

Organism UniProt Comment Textmining
Vibrio harveyi
-
-
-

Purification (Commentary)

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

Reaction

Reaction Comment Organism Reaction ID
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]

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

Substrates and Products (Substrate)

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

Subunits

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

Synonyms

Synonyms Comment Organism
luciferase
-
 Vibrio harveyi
Vibrio harveyi luciferase
-
 Vibrio harveyi

Temperature Optimum [°C]

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

pH Optimum

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

Cofactor

Cofactor Comment Organism Structure
FMNH2
-
 Vibrio harveyi