Cloned (Comment) | Organism |
---|---|
gene luxAB, expression of wild-type and mutant enzymes in Escherichia coli strain BL21 | Vibrio harveyi |
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 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 | 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 | UniProt | Comment | Textmining |
---|---|---|---|
Vibrio harveyi | - |
gene luxAB | - |
Purification (Comment) | Organism |
---|---|
recombinant wild-type and mutant enzymes from Escherichia coli strain BL21 | Vibrio harveyi |
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 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 | 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 | Comment | Organism |
---|---|---|
dimer | heterodimer | Vibrio harveyi |
Synonyms | Comment | Organism |
---|---|---|
bacterial luciferase | - |
Vibrio harveyi |
luciferase | - |
Vibrio harveyi |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
23 | - |
assay at | Vibrio harveyi |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
7 | - |
assay at | Vibrio harveyi |
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 |