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Literature summary for 1.14.14.3 extracted from

  • Low, J.C.; Tu, S.C.
    Functional roles of conserved residues in the unstructured loop of Vibrio harveyi bacterial luciferase (2002), Biochemistry, 41, 1724-1731.
    View publication on PubMed
Search for more literature for 1.14.14.3

Cloned(Commentary)

Cloned (Comment) Organism
gene luxA, expression of wild-type and mutant enzymes in Escherichia coli strain JM109 Vibrio harveyi

Protein Variants

Protein Variants Comment Organism
F261A site-directed mutagenesis, residue of the alpha-subunit, 0.19% of the wild-type activity, the bulky and hydrophobic nature of the alphaF261 residue is critical for activity Vibrio harveyi
F261D site-directed mutagenesis, residue of the alpha-subunit, 0.004% of the wild-type activity, the bulky and hydrophobic nature of the alphaF261 residue is critical for activity Vibrio harveyi
F261S site-directed mutagenesis, residue of the alpha-subunit, 0.13% of the wild-type activity, the bulky and hydrophobic nature of the alphaF261 residue is critical for activity Vibrio harveyi
F261Y site-directed mutagenesis, residue of the alpha-subunit, 2-3% of the wild-type activity, the bulky and hydrophobic nature of the alphaF261 residue is critical for activity Vibrio harveyi
G275A site-directed mutagenesis, residue of the alpha-subunit, 27% of the wild-type activity, the torsional flexibility of the alphaG275 residue is critical for activity Vibrio harveyi
G275F site-directed mutagenesis, residue of the alpha-subunit, 6-7% of the wild-type activity, the torsional flexibility of the alphaG275 residue is critical for activity Vibrio harveyi
G275I site-directed mutagenesis, residue of the alpha-subunit, 15% of the wild-type activity, the torsional flexibility of the alphaG275 residue is critical for activity Vibrio harveyi
G275P site-directed mutagenesis, residue of the alpha-subunit, 0.04% of the wild-type activity, the torsional flexibility of the alphaG275 residue is critical for activity Vibrio harveyi
G284P site-directed mutagenesis, residue of the alpha-subunit, 1-2% of the wild-type activity Vibrio harveyi

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
additional information
-
 additional information stopped flow spectroscopy Vibrio harveyi
0.0003
-
 FMNH2 recombinant wild-type enzyme, pH 7.0, 25°C Vibrio harveyi
0.0004
-
 decanal recombinant mutant alphaF261S, pH 7.0, 25°C Vibrio harveyi
0.0004
-
 decanal recombinant mutant alphaG275I, pH 7.0, 25°C Vibrio harveyi
0.0006
-
 FMNH2 recombinant mutant alphaG284P, pH 7.0, 25°C Vibrio harveyi
0.001
-
 decanal recombinant wild-type enzyme, pH 7.0, 25°C Vibrio harveyi
0.0016
-
 decanal recombinant mutant alphaF261Y, pH 7.0, 25°C Vibrio harveyi
0.0017
-
 decanal recombinant mutant alphaF261A, pH 7.0, 25°C Vibrio harveyi
0.0018
-
 decanal recombinant mutant alphaG275P, pH 7.0, 25°C Vibrio harveyi
0.0022
-
 decanal recombinant mutant alphaG275A, pH 7.0, 25°C Vibrio harveyi
0.0022
-
 FMNH2 recombinant mutant alphaG275P, pH 7.0, 25°C Vibrio harveyi
0.0027
-
 decanal recombinant mutant alphaG275F, pH 7.0, 25°C Vibrio harveyi
0.0037
-
 decanal recombinant mutant alphaG284P, pH 7.0, 25°C Vibrio harveyi
0.0046
-
 decanal recombinant mutant alphaF261D, pH 7.0, 25°C Vibrio harveyi
0.0075
-
 FMNH2 recombinant mutant alphaF261D, pH 7.0, 25°C Vibrio harveyi
0.0192
-
 FMNH2 recombinant mutant alphaF261Y, pH 7.0, 25°C Vibrio harveyi
0.0281
-
 FMNH2 recombinant mutant alphaF261A, pH 7.0, 25°C Vibrio harveyi
0.0358
-
 FMNH2 recombinant mutant alphaG275A, pH 7.0, 25°C Vibrio harveyi
0.0369
-
 FMNH2 recombinant mutant alphaF261S, pH 7.0, 25°C Vibrio harveyi
0.0411
-
 FMNH2 recombinant mutant alphaG275F, pH 7.0, 25°C Vibrio harveyi
0.0584
-
 FMNH2 recombinant mutant alphaG275I, pH 7.0, 25°C Vibrio harveyi

Organism

Organism UniProt Comment Textmining
Vibrio harveyi
-
-
-

Purification (Commentary)

Purification (Comment) Organism
recombinant wild-type and mutant enzymes from Escherichia coli strain JM109 to over 85% homogeneity Vibrio harveyi

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
decanal + FMNH2 + O2 formation of a 4a-hydroperoxy-FMN intermediate II Vibrio harveyi decanoate + FMN + H2O + hn
-
 ir
RCHO + FMNH2 + O2 formation of a 4a-hydroperoxy-FMN intermediate II Vibrio harveyi RCOOH + FMN + H2O + hn
-
 ir

Subunits

Subunits Comment Organism
More functional roles of conserved residues in the protease-labile, unstructured loop of the alpha-subunit, formed by residues 257-291, the loop undergoed conformational changes during catalysis, overview Vibrio harveyi

Synonyms

Synonyms Comment Organism
bacterial luciferase
-
 Vibrio harveyi

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
25
-
 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