Cloned (Comment) | Organism |
---|---|
ligated into pET21-b vector, expressed from pZCH2 in Escherichia coli BL21 (lambdaDE3) cell line | Vibrio harveyi |
Protein Variants | Comment | Organism |
---|---|---|
D262A | 90% reduced activity with octanal, 36% reduced activity with decanal, activity with dodecanal as the wild-type | Vibrio harveyi |
D265A | activity with octanal as the wild-type, 81% reduced activity with decanal, complete loss of dodecanal activity | Vibrio harveyi |
D271A | complete loss of octanal and decanal activity, 18% reduced activity with dodecanal | Vibrio harveyi |
H285A | 26% reduced activity with octanal, 74% reduced activity with decanal, complete loss of dodecanal activity | Vibrio harveyi |
K274A | 89% reduced activity with octanal, 21% reduced activity with decanal, 81% reduced activity with dodecanal | Vibrio harveyi |
K283A | complete loss of octanal and decanal activity, 96% reduced activity with dodecanal, does not significantly impede binding of decanal, results in destabilization of intermediate II, results in a loss in quantum yield comparable with that of the loop deletion mutant, binds reduced flavin more weakly | Vibrio harveyi |
K286A | 92% reduced activity with octanal, complete loss of decanal activity, 87% reduced activity with dodecanal, does not significantly impede binding of decanal, increase in exposure of reaction intermediates to a dynamic quencher, results in a loss in quantum yield comparable with that of the loop deletion mutant, binds reduced flavin more weakly | Vibrio harveyi |
R291A | 77% reduced activity with octanal, 58% reduced activity with decanal, 71% reduced activity with dodecanal | Vibrio harveyi |
W277A | 11% reduced activity with octanal, 50% reduced activity with decanal and dodecanal | Vibrio harveyi |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.008 | - |
decanal | wild-type | Vibrio harveyi | |
0.01 | - |
decanal | mutant K286A | Vibrio harveyi | |
0.0105 | - |
decanal | mutant K283A | Vibrio harveyi |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Vibrio harveyi | - |
- |
- |
Purification (Comment) | Organism |
---|---|
on nickel affinity column, to more than 90% purity | Vibrio harveyi |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
decanal + FMNH + O2 | - |
Vibrio harveyi | decanoic acid + FMN + H2O + light | - |
? | |
dodecanal + FMNH + O2 | - |
Vibrio harveyi | dodecanoic acid + FMN + H2O + light | - |
? | |
octanal + FMNH + O2 | - |
Vibrio harveyi | octanoic acid + FMN + H2O + light | - |
? |
Synonyms | Comment | Organism |
---|---|---|
bacterial luciferase | - |
Vibrio harveyi |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
FMN | - |
Vibrio harveyi |