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Literature summary for 1.14.14.3 extracted from

  • Lin, L.Y.C.; Szittner, R.; Friedman, R.; Meighen, E.A.
    Changes in the kinetics and emission spectrum on mutation of the chromophore-binding platform in Vibrio harveyi luciferase (2004), Biochemistry, 43, 3183-3194.
    View publication on PubMed

Cloned(Commentary)

Cloned (Comment) Organism
gene luxAB, expression of wild-type and mutant enzymes in Escherichia coli strain BL21 Vibrio harveyi

Protein Variants

Protein Variants Comment Organism
A75G/C106V/V173A site-directed mutagenesis, alpha-subunit residues, reduced activity compared to the wild-type enzyme, further red shift of emission spectrum Vibrio harveyi
A75G/C106V/V173C site-directed mutagenesis, alpha-subunit residues, reduced activity compared to the wild-type enzyme, further red shift of emission spectrum Vibrio harveyi
A75G/C106V/V173S site-directed mutagenesis, alpha-subunit residues, reduced activity compared to the wild-type enzyme, further red shift of emission spectrum Vibrio harveyi
A75G/C106V/V173T site-directed mutagenesis, alpha-subunit residues, reduced activity compared to the wild-type enzyme, further red shift of emission spectrum Vibrio harveyi
additional information substrate specificities of mutant enzymes and wild-type enzyme, overview, changes in the kinetics and emission spectrum on mutation of the chromophore-binding platform Vibrio harveyi
V173A site-directed mutagenesis, alpha-subunit residue, reduced activity and decreased stability of the C4a-hydroperoxyflavin intermediate compared to the wild-type enzyme, red shift of emission spectrum Vibrio harveyi
V173C site-directed mutagenesis, alpha-subunit residue, reduced activity and decreased stability of the C4a-hydroperoxyflavin intermediate compared to the wild-type enzyme, red shift of emission spectrum Vibrio harveyi
V173F site-directed mutagenesis, alpha-subunit residue, reduced activity and decreased stability of the C4a-hydroperoxyflavin intermediate compared to the wild-type enzyme, red shift of emission spectrum Vibrio harveyi
V173H site-directed mutagenesis, alpha-subunit residue, reduced activity and decreased stability of the C4a-hydroperoxyflavin intermediate compared to the wild-type enzyme, red shift of emission spectrum Vibrio harveyi
V173I site-directed mutagenesis, alpha-subunit residue, reduced activity and decreased stability of the C4a-hydroperoxyflavin intermediate compared to the wild-type enzyme, red shift of emission spectrum Vibrio harveyi
V173L site-directed mutagenesis, alpha-subunit residue, reduced activity and decreased stability of the C4a-hydroperoxyflavin intermediate compared to the wild-type enzyme, red shift of emission spectrum Vibrio harveyi
V173N site-directed mutagenesis, alpha-subunit residue, reduced activity and decreased stability of the C4a-hydroperoxyflavin intermediate compared to the wild-type enzyme, red shift of emission spectrum Vibrio harveyi
V173S site-directed mutagenesis, alpha-subunit residue, reduced activity and decreased stability of the C4a-hydroperoxyflavin intermediate compared to the wild-type enzyme, red shift of emission spectrum Vibrio harveyi
V173T site-directed mutagenesis, alpha-subunit residue, reduced activity and decreased stability of the C4a-hydroperoxyflavin intermediate compared to the wild-type enzyme, red shift of emission spectrum Vibrio harveyi

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
additional information
-
additional information affinity and dissociation constants for FMNH2 of wild-type and mutants enzymes, kinetics Vibrio harveyi

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
fatty aldehyde + FMNH2 + O2 Vibrio harveyi
-
fatty acid + FMN + H2O + hn
-
ir
additional information Vibrio harveyi luminescence pathway, overview ?
-
?

Organism

Organism UniProt Comment Textmining
Vibrio harveyi
-
gene luxAB
-

Purification (Commentary)

Purification (Comment) Organism
recombinant wild-type and mutant enzymes from Escherichia coli strain BL21 Vibrio harveyi

Reaction

Reaction Comment Organism Reaction ID
a long-chain aldehyde + FMNH2 + O2 = a long-chain fatty acid + FMN + H2O + hnu residue Val173 of the alpha-subunit is essential for flavin binding and located in the active site, formation of a C4a-hydroperoxyflavin intermediate Vibrio harveyi

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
additional information
-
luminescence activity and decay of wild-type and mutants with different aldehyde substrates Vibrio harveyi

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
decanal + FMNH2 + O2
-
Vibrio harveyi decanoate + FMN + H2O + hn
-
ir
dodecanal + FMNH2 + O2
-
Vibrio harveyi dodecanoate + FMN + H2O + hn
-
ir
fatty aldehyde + FMNH2 + O2
-
Vibrio harveyi fatty acid + FMN + H2O + hn
-
ir
additional information luminescence pathway, overview Vibrio harveyi ?
-
?
additional information substrate specificities of mutant enzymes and wild-type enzyme, overview Vibrio harveyi ?
-
?
nonanal + FMNH2 + O2
-
Vibrio harveyi nonanoate + FMN + H2O + hn
-
ir
octanal + FMNH2 + O2
-
Vibrio harveyi octanoate + FMN + H2O + hn
-
ir
RCHO + FMNH2 + O2 formation of a C4a-hydroperoxyflavin intermediate Vibrio harveyi RCOOH + FMN + H2O + hn
-
ir
undecanal + FMNH2 + O2
-
Vibrio harveyi undecanoate + FMN + H2O + hn
-
ir

Subunits

Subunits Comment Organism
dimer heterodimer Vibrio harveyi

Synonyms

Synonyms Comment Organism
bacterial luciferase
-
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 enzyme is complexed with flavin mononucleotide, enzyme possesses a binding platform for the isoalloxazine ring of flavin, a chromophore whose 7,8-dimethyl benzene plane interacts with the isopropyl chain of alphaVal173, structure-fuction analysis Vibrio harveyi