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Literature summary for 1.14.14.3 extracted from

  • Li, C.H.; Tu, S.C.
    Probing the functionalities of alphaGlu328 and alphaAla74 of Vibrio harveyi luciferase by site-directed mutagenesis and chemical rescue (2005), Biochemistry, 44, 13866-13873.
    View publication on PubMed
Search for more literature for 1.14.14.3

Activating Compound

Activating Compound Comment Organism Structure
Sodium acetate activates mutant E328A Vibrio harveyi

Protein Variants

Protein Variants Comment Organism
A74F site-directed mutagenesis, the mutant shows reduced activity and increased Km compared to the wild-type enzyme Vibrio harveyi
A74G site-directed mutagenesis, the mutant shows increased activity compared to the wild-type enzyme Vibrio harveyi
E328A site-directed mutagenesis, the mutant shows highly reduced activity and increased Km compared to the wild-type enzyme, the activity is rescued by addition of sodium acetate, but not by phosphate, at pH 6.0-8.0 with increasing activity at lower pH Vibrio harveyi
E328D site-directed mutagenesis, the mutant shows highly reduced activity and increased Km compared to the wild-type enzyme Vibrio harveyi
E328F site-directed mutagenesis, the mutant shows reduced activity and increased Km compared to the wild-type enzyme Vibrio harveyi
E328H site-directed mutagenesis, the mutant shows highly reduced activity and increased Km compared to the wild-type enzyme Vibrio harveyi
E328L site-directed mutagenesis, the mutant shows highly reduced activity and increased Km compared to the wild-type enzyme Vibrio harveyi
E328Q site-directed mutagenesis, the mutant shows highly reduced activity and increased Km compared 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 and Michaelis-Menten kinetics of wild-type and mutant enzymes Vibrio harveyi
0.0002
-
 FMNH2 pH 7.0, 23°C, recombinant mutants E328Q and E328H Vibrio harveyi
0.0003
-
 FMNH2 pH 7.0, 23°C, recombinant mutants E328L and E328A Vibrio harveyi
0.0004
-
 FMNH2 pH 7.0, 23°C, recombinant mutant E328D Vibrio harveyi
0.0006
-
 FMNH2 pH 7.0, 23°C, recombinant wild-type enzyme Vibrio harveyi
0.0015
-
 FMNH2 pH 7.0, 23°C, recombinant mutant A74G Vibrio harveyi
0.0016
-
 decanal pH 7.0, 23°C, recombinant wild-type enzyme and mutant A74G Vibrio harveyi
0.0023
-
 decanal pH 7.0, 23°C, recombinant mutant E328Q Vibrio harveyi
0.0033
-
 decanal pH 7.0, 23°C, recombinant mutant E328A Vibrio harveyi
0.0039
-
 FMNH2 pH 7.0, 23°C, recombinant mutant E328F Vibrio harveyi
0.0045
-
 decanal pH 7.0, 23°C, recombinant mutant E328F Vibrio harveyi
0.005
-
 decanal pH 7.0, 23°C, recombinant mutant E328D Vibrio harveyi
0.0087
-
 FMNH2 pH 7.0, 23°C, recombinant mutant A74F Vibrio harveyi
0.0095
-
 decanal pH 7.0, 23°C, recombinant mutant E328H Vibrio harveyi
0.0097
-
 decanal pH 7.0, 23°C, recombinant mutant E328L Vibrio harveyi
0.0173
-
 decanal pH 7.0, 23°C, recombinant mutant A74F Vibrio harveyi

Organism

Organism UniProt Comment Textmining
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, alphaE238, alphaA75, and alphaA74 are catalytically important active site residues of the alpha-subunit, structure-function relationship Vibrio harveyi
a

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
additional information
-
 light and dark 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

Subunits

Subunits Comment Organism
dimer alphabeta heterodimer, structure-function relationship, overview 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