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Information on EC 1.14.14.3 - bacterial luciferase and Organism(s) Vibrio harveyi and UniProt Accession P07740
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1.14.14.3 bacterial luciferaseIUBMB Comments
The reaction sequence starts with the incorporation of a molecule of oxygen into reduced FMN bound to the enzyme, forming luciferase peroxyflavin. The peroxyflavin interacts with an aliphatic long-chain aldehyde, producing a highly fluorescent species believed to be luciferase hydroxyflavin. The enzyme is highly specific for reduced FMN and for long-chain aliphatic aldehydes with eight carbons or more. The highest efficiency is achieved with tetradecanal. cf. EC 1.13.12.18, dinoflagellate luciferase.
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The taxonomic range for the selected organisms is: Vibrio harveyi
The expected taxonomic range for this enzyme is: Bacteria, Archaea, Eukaryota
The expected taxonomic range for this enzyme is: Bacteria, Archaea, Eukaryota
Synonyms
luciferase, bacterial luciferase, luxab, luxcdabe, vibrio harveyi luciferase, vibrio fischeri luciferase, aldehyde monooxygenase, gluc luciferase, 
more
topSYNONYM 
ORGANISM
UNIPROT
COMMENTARY 
LITERATURE
aldehyde monooxygenase
-
-
-
-
alkanal monooxygenase (FMN)
-
-
-
-
bacterial luciferase
luciferase
LuxAB
Vibrio fischeri luciferase
-
-
-
-
bacterial luciferase
-
-
-
-
bacterial luciferase
-
-
luciferase
-
-
-
-
luciferase
-
-
LuxAB
monocistronic gene that encodes monomeric bacterial luciferase
REACTION
REACTION DIAGRAM
COMMENTARY
ORGANISM
UNIPROT
LITERATURE
a long-chain aldehyde + FMNH2 + O2 = a long-chain fatty acid + FMN + H2O + hnu
reaction mechanism and scheme, cofactor binding
-
a long-chain aldehyde + FMNH2 + O2 = a long-chain fatty acid + FMN + H2O + hnu
reaction mechanism, Cys106 is essential for stabilization of the reaction intermediate, for consumption of aldehyde substrate, and for activity
-
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
-
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
-
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, overview, catalytically important active site residues of the alpha-subunit are alphaF46, alphaF49, alphaF114, and alphaF117, alphaF46 and alphaF117 also appear to be involved in the binding of reduced flavin substrate
-
REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
redox reaction
-
-
-
-
oxidation
-
-
-
-
reduction
-
-
-
-
PATHWAY SOURCE
PATHWAYS
-
-
SYSTEMATIC NAME
IUBMB Comments
long-chain-aldehyde,FMNH2:oxygen oxidoreductase (1-hydroxylating, luminescing)
The reaction sequence starts with the incorporation of a molecule of oxygen into reduced FMN bound to the enzyme, forming luciferase peroxyflavin. The peroxyflavin interacts with an aliphatic long-chain aldehyde, producing a highly fluorescent species believed to be luciferase hydroxyflavin. The enzyme is highly specific for reduced FMN and for long-chain aliphatic aldehydes with eight carbons or more. The highest efficiency is achieved with tetradecanal. cf. EC 1.13.12.18, dinoflagellate luciferase.
CAS REGISTRY NUMBER
COMMENTARY
9014-00-0
-
SUBSTRATE
PRODUCT
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate)
(Substrate)
LITERATURE
(Substrate)
(Substrate)
COMMENTARY
(Product)
(Product)
LITERATURE
(Product)
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
r=reversible
ir=irreversible
?=not specified
(E)-tetradec-2-enal + FMNH2 + O2
(E)-tetradec-2-enoate + FMN + H2O + hn
-
-
-
-
?
4-N,N-(dimethyl)aminonaphthalene-9-N-(11-aldehydedodecyl)-1,8-dicarboximide + FMNH2 + O2
? + FMN + H2O + hnu
-
-
-
-
?
4-N,N-(dimethyl)aminonaphthalene-9-N-(9-aldehyde-decyl)-1,8-dicarboximide + FMNH2 + O2
? + FMN + H2O + hnu
-
-
-
-
?
4-N-(11-aldehyde-dodecyl)-7-N,N-dimethylsulfonic-2,1,3-benzoxadiazole + FMNH2 + O2
? + FMN + H2O + hnu
-
-
-
-
?
4-N-(9-aldehyde-decyl)-7-N,N-dimethylsulfonic-2,1,3-benzoxadiazole + FMNH2 + O2
? + FMN + H2O + hnu
-
-
-
-
?
a long-chain aldehyde + FMNH2 + O2
a long-chain fatty acid + FMN + H2O + hv
-
-
-
?
decanal + riboflavin + O2
?
-
riboflavin is a very poor substrate for bacterial luciferase
-
-
?
an aldehyde + FMNH2 + O2
a carboxylate + FMN + H2O + hnu
-
-
-
-
?
an aldehyde + FMNH2 + O2
a carboxylate + FMN + H2O + hnu
-
a long chain aliphatic aldehyde as substrate
-
-
?
decanal + FMNH + O2
decanoic acid + FMN + H2O + light
-
-
348545, 348546, 348547, 348548, 348549, 348550, 348551, 348552, 348553, 348554, 348555, 348557, 348558, 348559, 348560, 348562, 348564, 348565, 348566, 348567, 348568, 348569, 348570, 348571, 348572, 348573, 348574, 348575, 348576, 348577, 348579, 348581, 348582, 348583, 348584, 348585, 348587, 348588, 348589, 348597, 348599, 348600, 348601, 348602, 348604, 348607, 348608
-
-
ir
decanal + FMNH2 + O2
decanoate + FMN + H2O + hn
-
formation of a 4a-hydroperoxy-FMN intermediate II
-
-
ir
decanal + FMNH2 + O2
decanoic acid + FMN + H2O + hv
-
-
light emission at 490 nm
-
?
dodecanal + FMNH + O2
dodecanoic acid + FMN + H2O + light
-
-
348545, 348546, 348547, 348548, 348549, 348550, 348551, 348552, 348553, 348554, 348555, 348557, 348558, 348559, 348560, 348562, 348564, 348565, 348566, 348567, 348568, 348569, 348570, 348571, 348572, 348573, 348574, 348575, 348576, 348577, 348579, 348581, 348582, 348583, 348584, 348585, 348587, 348588, 348589, 348597, 348599, 348600, 348601, 348602, 348604, 348607, 348608
-
-
ir
octanal + FMNH + O2
octanoic acid + FMN + H2O + light
-
-
348545, 348546, 348547, 348548, 348549, 348550, 348551, 348552, 348553, 348554, 348555, 348557, 348558, 348559, 348560, 348562, 348564, 348565, 348566, 348567, 348568, 348569, 348570, 348571, 348572, 348573, 348574, 348575, 348576, 348577, 348579, 348581, 348582, 348583, 348584, 348585, 348587, 348588, 348589, 348597, 348599, 348600, 348601, 348602, 348604, 348607, 348608
-
-
ir
RCHO + FMNH2 + O2
RCOOH + FMN + H2O + hn
-
formation of a 4a-hydroperoxy-FMN intermediate II
-
-
ir
RCHO + FMNH2 + O2
RCOOH + FMN + H2O + hn
-
formation of a C4a-hydroperoxyflavin intermediate
-
-
ir
RCHO + FMNH2 + O2
RCOOH + FMN + H2O + hnu
-
reduced FMN, i.e. FMNH2, generated by several species of flavin reductases, is utilized along with a long-chain aliphatic aldehyde and molecular oxygen by luciferase as substrates for the bioluminescence reaction, direct transfer of reduced flavin cofactor and reduced flavin product of reductase to luciferase, NADPH-specific FMN reductase and luciferase form a complex in vivo, reduction of reductase-bound FMN cofactor by NADPH is reversible, allowing the cellular contents of NADP+ and NADPH as a factor for the regulation of the production of FMNH2 by FRPVh for luciferase bioluminescence, overview
-
-
?
additional information
?
-
-
aldehydes of chain-length 8 or more required
-
-
?
additional information
?
-
-
complex formation in a 1:1 molar ratio between monomeric, but not dimeric, NADPH:FMN oxidoreductase FRP and luciferase for direct transfer of cofactor FMNH2
-
-
?
additional information
?
-
-
the enzyme plays a role in protection of cells against oxidative stress
-
-
?
additional information
?
-
-
substrate specificities of mutant enzymes and wild-type enzyme, overview
-
-
?
additional information
?
-
-
Vibrio harveyi NADPH-specific flavin reductase FRP transfers reduced riboflavin-5'-phosphate to luciferase by both free diffusion and direct transfer, resulting inbioluminescence production, FRP:luciferase coupled bioluminescence reaction, overview, increases in oxygen concentration lead to gradual decreases of the peak bioluminescence intensity, Km for FMN, and Km for NADPH of NADPH-specific flavin reductase in the coupled reaction with luciferase
-
-
?
additional information
?
-
-
active site hydrophobicity is critical to the bioluminescence activity of Vibrio harveyi luciferase
-
-
?
additional information
?
-
-
the 4a-hydroperoxy-4a,5-dihydroFMN intermediate luciferase transforms from a low quantum yield IIx to a high quantum yield IIy fluorescent species on exposure to excitation light
-
-
?
additional information
?
-
-
FMNH2 binds to a mobile loop of 29 amino acids in the luciferase protein, loop modeling of ligand-free and -bound enzyme, conformation and dynamics, overview
-
-
?
additional information
?
-
-
enzyme accepts unsaturated aldehydes as substrates but light emission drops drastically compared to saturated aldehydes. The onset and the decay rate of bioluminescence are much slower, when using unsaturated substrates. As a result the duration of the light emission is doubled
-
-
?
NATURAL SUBSTRATE
NATURAL PRODUCT
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate)
(Substrate)
LITERATURE
(Substrate)
(Substrate)
COMMENTARY
(Product)
(Product)
LITERATURE
(Product)
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
r=reversible
ir=irreversible
?=not specified
an aldehyde + FMNH2 + O2
a carboxylate + FMN + H2O + hnu
-
-
-
-
?
decanal + FMNH + O2
decanoic acid + FMN + H2O + light
-
-
348545, 348546, 348547, 348548, 348549, 348550, 348551, 348552, 348553, 348554, 348555, 348557, 348558, 348559, 348560, 348562, 348564, 348565, 348566, 348567, 348568, 348569, 348570, 348571, 348572, 348573, 348574, 348575, 348576, 348577, 348579, 348581, 348582, 348583, 348584, 348585, 348587, 348588, 348589, 348597, 348599, 348600, 348601
-
-
ir
RCHO + FMNH2 + O2
RCOOH + FMN + H2O + hnu
-
reduced FMN, i.e. FMNH2, generated by several species of flavin reductases, is utilized along with a long-chain aliphatic aldehyde and molecular oxygen by luciferase as substrates for the bioluminescence reaction, direct transfer of reduced flavin cofactor and reduced flavin product of reductase to luciferase, NADPH-specific FMN reductase and luciferase form a complex in vivo, reduction of reductase-bound FMN cofactor by NADPH is reversible, allowing the cellular contents of NADP+ and NADPH as a factor for the regulation of the production of FMNH2 by FRPVh for luciferase bioluminescence, overview
-
-
?
additional information
?
-
-
complex formation in a 1:1 molar ratio between monomeric, but not dimeric, NADPH:FMN oxidoreductase FRP and luciferase for direct transfer of cofactor FMNH2
-
-
?
additional information
?
-
-
the enzyme plays a role in protection of cells against oxidative stress
-
-
?
additional information
?
-
-
Vibrio harveyi NADPH-specific flavin reductase FRP transfers reduced riboflavin-5'-phosphate to luciferase by both free diffusion and direct transfer, resulting inbioluminescence production, FRP:luciferase coupled bioluminescence reaction, overview, increases in oxygen concentration lead to gradual decreases of the peak bioluminescence intensity, Km for FMN, and Km for NADPH of NADPH-specific flavin reductase in the coupled reaction with luciferase
-
-
?
COFACTOR
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
4a-hydroxy-4a,5-dihydroriboflavin-5'-phosphate
-
model bioluminescence emitter molecule, binding and fluorescence quantum yield studies, complexed with the enzyme in a 1:1 molcular ratio
additional information
-
the 4a-hydroperoxy-4a,5-dihydroFMN intermediate luciferase transforms from a low quantum yield IIx to a high quantum yield IIy fluorescent species on exposure to excitation light
-
FMNH2
-
-
348552, 348579, 657751, 657923, 657927, 657939, 657994, 658495, 671610, 671978, 671985, 698992, 711341
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
FMNH2
-
reduced FMN, i.e. FMNH2, generated by several species of flavin reductases, is utilized along with a long-chain aliphatic aldehyde and molecular oxygen by luciferase as substrates for the bioluminescence reaction, direct transfer of reduced flavin cofactor and reduced flavin product of reductase to luciferase, overview
FMNH2
-
binds to a mobile loop of 29 amino acids in the luciferase protein, structure and conformation, overview
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
5-decyl-4a-hydroxy-4a,5-dihydroriboflavin-5'-phosphate
-
binding and fluorescence quantum yield studies of the substance as a model, complexed with the enzyme in a 1:1 molcular ratio leading to 80% and 90% inhibition of wild-type and mutant C106A at 0.01 mM, respectively, binds to the active site
potassium iodide
-
quenches the fluorescence of FMN effectively at 0.2 M, and enhances the decay of wild-type and HFOOH enzymes, the wild-type enzyme forms an inactive complex with KI
8-Anilino-1-naphthalenesulfonate
-
inhibitor binding site separate from FMN-binding site by 30 A
n-decanal
-
reversible substrate inhibition, depending on phosphate concentration
additional information
-
luxA mutant and luxB mutant strains are more sensitive to oxidants like H2O2, cumene hydroperoxide, tert-butyl hydroperoxide, or ferrous sulfate, than the wild-type strain, growth behaviour overview
-
additional information
-
increases in oxygen concentration lead to gradual decreases of the peak bioluminescence intensity, Km for FMN, and Km for NADPH of NADPH-specific flavin reductase in the coupled reaction with luciferase
-
ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
0.26
4-N,N-(dimethyl)aminonaphthalene-9-N-(11-aldehydedodecyl)-1,8-dicarboximide
-
pH 7.0, temperature not specified in the publication
0.72
4-N,N-(dimethyl)aminonaphthalene-9-N-(9-aldehyde-decyl)-1,8-dicarboximide
-
pH 7.0, temperature not specified in the publication
0.72
4-N-(11-aldehyde-dodecyl)-7-N,N-dimethylsulfonic-2,1,3-benzoxadiazole
-
pH 7.0, temperature not specified in the publication
1.79
4-N-(9-aldehyde-decyl)-7-N,N-dimethylsulfonic-2,1,3-benzoxadiazole
-
pH 7.0, temperature not specified in the publication
0.0009
FMN
-
in the presence of different flavin concentrations, 0.001 mM Fre oxidoreductase, 10 mM decanal, and 0.01 mM NADPH and 0.005 mM luciferase
0.0013
riboflavin
-
in the presence of different flavin concentrations, 0.001 mM Fre oxidoreductase, 10 mM decanal, and 0.01 mM NADPH and 0.005 mM luciferase
0.00113
decanal
-
recombinant wild type enzyme, at 22°C, pH not specified in the publication
0.00165
decanal
-
luciferase-mOrange fusion enzyme, at 22°C, pH not specified in the publication
0.00018
FMNH2
-
luciferase-mOrange fusion enzyme, at 22°C, pH not specified in the publication
0.0002
FMNH2
-
recombinant wild type enzyme, at 22°C, pH not specified in the publication
0.0003
FMNH2
-
pH 7.0, 23°C, recombinant mutants F46Y, F114A, F117Y, and F114Y
additional information
additional information
-
affinity and dissociation constants for FMNH2 of wild-type and mutants enzymes, kinetics
-
additional information
additional information
-
detailed reaction and folding kinetics, thermodynamics
-
additional information
additional information
-
enzyme activities in complex formation, kinetics, dissociation constants
-
additional information
additional information
-
kinetics, substrate and cofactor binding
-
additional information
additional information
-
kinetics of the FRP:luciferase coupled bioluminescence reaction
-
additional information
additional information
-
stopped-flow and Michaelis-Menten kinetics of wild-type and mutant enzymes
-
additional information
additional information
-
stopped-flow kinetics of wild-type and mutant enzymes
-
additional information
additional information
-
kinetics of FMN reductase-luciferase complex formation, overview
-
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
additional information
additional information
-
single turnover, luminescence
-
IC50 VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
0.03
8-Anilino-1-naphthalenesulfonate
Vibrio harveyi
-
in 50 mM phosphate buffer, pH 7.0 at 23°C
0.095
N-phenacylthiazolium bromide
Vibrio harveyi
-
in 50 mM phosphate buffer, pH 7.0 at 23°C
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
additional information
additional information
-
luminescence activity and decay of wild-type and mutants with different aldehyde substrates
additional information
-
light and dark quantum yield of wild-type and mutant enzymes, overview
additional information
-
quantum yield of wild-type and mutant enzymes, overview
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
ORGANISM
COMMENTARY
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY
GeneOntology No.
LITERATURE
SOURCE
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
metabolism
physiological function
metabolism
during the reaction, the first observed flavin C4a intermediate at pH below 8.5 is the protonated flavin C4a-hydroperoxide, which loses a proton to become an active flavin C4a-peroxide. Residue His44 plays an essential role in deprotonating the flavin C4a-hydroperoxide and initiating chemical catalysis. Residue His45 has a role in binding reduced FMN
metabolism
the enzyme controls the formation of the FMNH-C4a-OO- intermediate via the conserved His44 residue, the steps are: H+ reacts with O2 to generate +OOH. +OOH attacks C4a of FMNH- to generate FMNH-C4a-OOH. H+ is transferred from FMNH-C4a?OOH to His44 to generate FMNH-C4a-OO- while His44 stabilizes FMNH-C4a-OO- by forming a hydrogen bond to an oxygen atom
physiological function
-
bacterial luciferase requires an exogenous source of reduced flavin mononucleotide for bioluminescence activity. There is no stable complex formation between luciferase and oxidoreductases Fre/NfsA from Escherichia coli or Frp from Vibrio harveyi, which are believed to provide reduced flavin for luciferase activity. No difference in the levels of luciferase expression in either the DELTAfre or DELTANsfA strains
physiological function
-
luciferase is the enzyme responsible for light emission in bioluminescent bacteria, it catalyzes the reaction of reduced flavin mononucleotide FMNH2, O2, and an aliphatic aldehyde to yield FMN, the corresponding carboxylic acid, and blue-green light
physiological function
-
the luxCDABE encoded protein products synergistically generate bioluminescent light signals exclusive of supplementary substrate additions or exogenous manipulations
UNIPROT
ENTRY NAME
ORGANISM
NO. OF AA
NO. OF TRANSM. HELICES
MOLECULAR WEIGHT[Da]
SOURCE
SEQUENCE
LOCALIZATION PREDICTION
The reliability class of the localization prediction ranges from 1 (very reliable) to 5 (not reliable).
The reliability class of the localization prediction ranges from 1 (very reliable) to 5 (not reliable). PDB
SCOP
CATH
UNIPROT
ORGANISM
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
77000 - 78000
SUBUNIT
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
dimer
heterodimer
additional information
dimer
-
alphabeta heterodimer containing a single active site at a cleft in the alpha subunit
additional information
-
analysis of subunit interface structure and role in the conformational stability of the heterodimeric enzyme, the beta-sunbunit can self-associate to form a stable but inactive homodimer, unfolding in a four-state mechanism
additional information
-
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
additional information
-
structure-function relationship, roles of Cys106, Ala75, Ala74, and the isoalloxazine ring of FMN
additional information
-
luciferase is composed of two homologous subunits designated alpha and beta, both of which assume the TIM barrel fold. Although the beta-subunit is required for activity, the catalytic site resides exclusively on the alpha-subunit. The most substantial compositional difference between subunits corresponds to a highly conserved stretch of residues between positions 260 and 290 unique to the alpha chains of luminous bacteria. In the luciferase/FMN complex, the asymmetric unit contains two beta/alpha-heterodimers
CRYSTALLIZATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
bacterial luciferase/FMN complex, by the hanging drop method, at 2.3 A resolution. Crystals of recombinant luciferase are grown at room temperature prior to soaking with millimolar concentrations of FMN. Belongs to space group P212121. The isoalloxazine ring is coordinated by an unusual cis-Ala-Ala peptide bond. The reactive sulfhydryl group of Cys106 projects toward position C-4a, the site of flavin oxygenation. Mobile loop that is crystallographically disordered, appears to be a boundary between solvent and the active center. Within this portion of the protein, there is a single contact between Phe272 of the R subunit and Tyr151 of the beta subunit
comparison of the conformational transitions of luciferases from Vibrio harveyi and Photobacterium leiognathi during equilibrium unfolding with urea. Vibrio harveyi luciferase in its native state demonstrates a higher fluorescence intensity per one protein molecule, but a shorter fluorescence lifetime, than Photobacterium leiognathi luciferase. During the first stage of denaturation (at more than 2 M of urea), for V. harveyi luciferase, the fluorescence lifetimes tau1 and tau2 show an increase, while for the P. leiognathi enzyme, the lifetime components decrease. This stage includes the unfolding of the C-terminal domain of the luciferase alpha-subunit. Subunit dissociation does not influence the optical characteristics of either of the luciferases. The unfolding of the subunits occurs in the same way for the two proteins
PROTEIN VARIANTS
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
Y151A
binds FMNH2 more weakly in comparison to the wild-type, substitution at position 151 on the beta subunit causes reductions in activity and total quantum yield
Y151D
binds FMNH2 more weakly in comparison to the wild-type, substitution at position 151 on the beta subunit causes reductions in activity and total quantum yield
Y151K
binds FMNH2 more weakly in comparison to the wild-type, substitution at position 151 on the beta subunit causes reductions in activity and total quantum yield
Y151R
binds FMNH2 more weakly in comparison to the wild-type, substitution at position 151 on the beta subunit causes reductions in activity and total quantum yield
Y151T
binds FMNH2 more weakly in comparison to the wild-type, substitution at position 151 on the beta subunit causes reductions in activity and total quantum yield
Y151W
least active mutant, binds reduced flavin with wild-type affinity, substitution at position 151 on the beta subunit causes reductions in activity and total quantum yield
A74F
-
site-directed mutagenesis, the mutant shows reduced activity and increased Km compared to the wild-type enzyme
A74G
-
site-directed mutagenesis, the mutant shows increased activity compared to the wild-type enzyme
A75G/C106V/V173A
-
site-directed mutagenesis, alpha-subunit residues, reduced activity compared to the wild-type enzyme, further red shift of emission spectrum
A75G/C106V/V173C
-
site-directed mutagenesis, alpha-subunit residues, reduced activity compared to the wild-type enzyme, further red shift of emission spectrum
A75G/C106V/V173S
-
site-directed mutagenesis, alpha-subunit residues, reduced activity compared to the wild-type enzyme, further red shift of emission spectrum
A75G/C106V/V173T
-
site-directed mutagenesis, alpha-subunit residues, reduced activity compared to the wild-type enzyme, further red shift of emission spectrum
A81H
-
site-directed mutagenesis, residue of the alpha-subunit, mutant shows 13% of wild-type activity
alphaF114A
-
site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme due to reduced hydrophobicity of the active site
alphaF114D
-
site-directed mutagenesis, the mutant shows highly reduced activity and increased Km compared to the wild-type enzyme due to reduced hydrophobicity of the active site
alphaF114S
-
site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme due to reduced hydrophobicity of the active site
alphaF114Y
-
site-directed mutagenesis, the mutant shows slightly reduced activity compared to the wild-type enzyme
alphaF117A
-
site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme due to reduced hydrophobicity of the active site
alphaF117D
-
site-directed mutagenesis, the mutant shows highly reduced activity and increased Km compared to the wild-type enzyme due to reduced hydrophobicity of the active site
alphaF117S
-
site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme due to reduced hydrophobicity of the active site
alphaF117Y
-
site-directed mutagenesis, the mutant shows slightly reduced activity compared to the wild-type enzyme
alphaF327A
-
site-directed mutagenesis, mutant activity is similar to the wild-type enzyme
alphaF46A
-
site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme due to reduced hydrophobicity of the active site
alphaF46D
-
site-directed mutagenesis, the mutant shows highly reduced activity and increased Km compared to the wild-type enzyme due to reduced hydrophobicity of the active site
alphaF46S
-
site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme due to reduced hydrophobicity of the active site
alphaF46Y
-
site-directed mutagenesis, the mutant shows slightly reduced activity compared to the wild-type enzyme
alphaF49A
-
site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme due to reduced hydrophobicity of the active site
alphaF49D
-
site-directed mutagenesis, the mutant shows highly reduced activity and increased Km compared to the wild-type enzyme due to reduced hydrophobicity of the active site
alphaF49S
-
site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme due to reduced hydrophobicity of the active site
alphaF49Y
-
site-directed mutagenesis, the mutant shows slightly reduced activity compared to the wild-type enzyme
alphaF6A
-
site-directed mutagenesis, mutant activity is similar to the wild-type enzyme
alphaH44A
C106A
-
site-directed mutagenesis, catalytic properties are similar to the wild-type enzyme, mutant shows 60% of wild-type quantum yield
C106V
-
site-directed mutagenesis, highly reduced ability to stabilize the reaction intermediate due to interaction between Val106 and Ala75 side chains, and therefore highly reduced activity and increased thermal lability compared to the wild-type enzyme
C106V/A75G
-
site-directed mutagenesis, mutation of Ala75 restores about 90% of the activity abolished by mutation of Cys106, shift in the light emission spectrum to that of Photobacterium phosphoreum possessing Val and Gly at positions 106 and 75, respectively
D262A
-
90% reduced activity with octanal, 36% reduced activity with decanal, activity with dodecanal as the wild-type
D265A
-
activity with octanal as the wild-type, 81% reduced activity with decanal, complete loss of dodecanal activity
D271A
-
complete loss of octanal and decanal activity, 18% reduced activity with dodecanal
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
E328D
-
site-directed mutagenesis, the mutant shows highly reduced activity and increased Km compared to the wild-type enzyme
E328F
-
site-directed mutagenesis, the mutant shows reduced activity and increased Km compared to the wild-type enzyme
E328H
-
site-directed mutagenesis, the mutant shows highly reduced activity and increased Km compared to the wild-type enzyme
E328L
-
site-directed mutagenesis, the mutant shows highly reduced activity and increased Km compared to the wild-type enzyme
E328Q
-
site-directed mutagenesis, the mutant shows highly reduced activity and increased Km compared to the wild-type enzyme
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
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
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
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
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
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
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
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
G284P
-
site-directed mutagenesis, residue of the alpha-subunit, 1-2% of the wild-type activity
H285A
-
26% reduced activity with octanal, 74% reduced activity with decanal, complete loss of dodecanal activity
H81A
-
site-directed mutagenesis, residue of the beta-subunit, mutant shows 59% of wild-type activity
H81A/E89D
-
site-directed mutagenesis, residues of the beta-subunit, mutant shows 13% of wild-type activity
H82A
-
site-directed mutagenesis, residue of the beta-subunit, mutant shows 22% of wild-type activity
K274A
-
89% reduced activity with octanal, 21% reduced activity with decanal, 81% reduced activity with dodecanal
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
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
R291A
-
77% reduced activity with octanal, 58% reduced activity with decanal, 71% reduced activity with dodecanal
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
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
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
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
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
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
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
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
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
W277A
-
11% reduced activity with octanal, 50% reduced activity with decanal and dodecanal
additional information
alphaH44A
-
rapid decay of the 4a-hydroperoxy-4a,5-dihydroFMN intermediate enzyme, HFOOH, in the mutant
additional information
-
Saccharomyces cerevisiae recombinantly expressing the Vibrio harveyi luciferase produces bright and stable luminescence, transformed yeast strains can grow on 0.5% v/v Z-9-tetradecenal, but die on 0.005% v/v decanal
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
additional information
-
immobilization of the FMN reductase-luciferase complex
additional information
-
codon optimization of the luxCDE and frp genes, e.g. adaptation of the bacterial protein to mammalian temperature of 37°C, detailed overview
pH STABILITY
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
37
wild-type enzyme has a 90% reduction in activity after 92 min. Comparable loss for the Y151W mutant after only 11 min
37
-
comparison of stability of wild-type enzyme and gene-fusion monomeric enzyme
40
-
wild-type enzyme and mutanbt A75G: loss of 60% activity within 50 min, mutants C106V and C106V/A75G show increased thermolability loosing 99% and 90% activity, respectively
GENERAL STABILITY
ORGANISM
UNIPROT
LITERATURE
STORAGE STABILITY
ORGANISM
UNIPROT
LITERATURE
-20°C, 50 mM potassium phosphate buffer, pH 7.0, protein concentration 1 mg/ml , -20°C, 0.1 M phosphate buffer, pH 7, 0.1 mM dithiothreitol, 1 mM EDTA
-
0-4°C, immobilized enzyme, 0.1 mM dithiothreitol, 20% loss of activity in 3 days
-
PURIFICATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
DEAE-Sepharose column chromatography, Superdex 200 gel filtration, and Mono Q column chromatography
-
recombinant wild-type and mutant C106A enzymes from Escherichia coli strain JM101 to over 95% purity
-
recombinant wild-type and mutant enzymes from Escherichia coli strain JM101 to homogeneity
-
recombinant wild-type and mutant enzymes from Escherichia coli strain JM109 to over 85% homogeneity
-
CLONED (Commentary)
ORGANISM
UNIPROT
LITERATURE
amplified from the pJHD500 plasmid and ligated into a pET21b vector, expressed from pZCH2 in an Escherichia coli BL21 (lambdaDE3) cell line after growth to an OD600 of 0.5
expressed in Escherichia coli from pJHD500, ligated into a pET21b vector. Luciferase subcloned from pZCH2 into a pASKIBA-3c vector with the restriction sites XbaI and XhoI. The resulting luciferase containing a strep-II tag on the C terminus of the beta-subunit (pZCB4) expressed
-
expressed in Escherichia coli JM109 (native enzyme) and BL21(DE3) cells (luciferase-mOrange fusion enzyme)
-
expression of fused luxA and luxB genes in Saccharomyces cerevisiae, Bacillus subtilis, plant cells, plasmid expression vector and in Escherichia coli
-
expression of seperated luxA and luxB gene in Escherichia coli JM109
-
expression of the enzyme in Mycobacterium tuberculosis under control of the inducible/repressible promotor of the alanine dehydrogenase from Mycobacterium tuberculosis strain H37Rv, usage of a mycobacterial-Escherichia coli shuttle vector
-
expression of wild-type and mutant C106A enzymes in Escherichia coli strain JM101
-
gene luxA, expression of wild-type and mutant enzymes in Escherichia coli strain JM109
-
gene luxAB, expression of wild-type and mutant enzymes in Escherichia coli strain BL21
-
gene luxAB, functional coexpression in Saccharomyces cerevisiae with NADPH-specific FMN reductase FRP from Vibrio harveyi, subcloning in Escherichia coli
-
ligated into pET21-b vector, expressed from pZCH2 in Escherichia coli BL21 (lambdaDE3) cell line
-
overexpression of wild-type and mutant enzymes in Escherichia coli strain JM101
-
the bacterial luciferase lux gene cassette consists of five genes, luxCDABE. The lux operon is re-synthesized through a process of multibicistronic, codon-optimization to demonstrate self-directed bioluminescence emission in a mammalian HEK-293 cell line in vitro and in vivo, overview. To overcome the limitations by FMNH2 supply, co-expression of a constitutively expressed flavin reductase gene frp from Vibrio harveyi is performed leading to a 151fold increased increase in bioluminescence in cells expressing mammalian codon-optimized luxCDE and frp genes
-
RENATURED/Commentary
ORGANISM
UNIPROT
LITERATURE
in the presence of the DnaKJE chaperone system thermally inactivated monomeric bacterial luciferase refolds. Monomeric bacterial luciferase thermally inactivated in the presence of ATPindependent trigger factor is not able to refold
APPLICATION
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
agriculture
-
engineering of broad-host-range Erwinia amylovora virus Y2 to enhance its killing activity and for use as a luciferase reporter phage. The reporter phage Y2::luxAB transduces bacterial luciferase into host cells and induces synthesis of large amounts of a LuxAB luciferase fusion. After the addition of aldehyde substrate, bioluminescence can be monitored, and enables rapid and specific detection of low numbers of viable bacteria
analysis
molecular biology
analysis
-
enzyme can be used to monitor changes in gene expression as a reporter system in slow-growing mycobacteria, i.e. Mycobacterium tuberculosis strain H37Ra, determination of recombinant enzyme decay rate
analysis
-
the enzyme is used as a reporter system tool for analysis of promoter and gene expression activity, overview
molecular biology
-
enzyme can be used to monitor changes in gene expression as a reporter system in slow-growing mycobacteria, i.e. Mycobacterium tuberculosis strain H37Ra, determination of recombinant enzyme decay rate
molecular biology
-
the enzyme is used as a reporter system tool for analysis of promoter and gene expression activity, overview
REF.
AUTHORS
TITLE
JOURNAL
VOL.
PAGES
YEAR
ORGANISM (UNIPROT)
PUBMED ID
SOURCE
Szittner, R.; Meighen, E.
Nucleotide sequence, expression, and properties of luciferase coded by lux genes from a terrestrial bacterium
J. Biol. Chem.
265
16581-16587
1990
Photorhabdus luminescens, Vibrio harveyi
Makemson, J.C.
A cyanide-aldehyde complex inhibits bacterial luciferase
J. Bacteriol.
172
4725-4727
1990
Vibrio harveyi
Curry, S.; Lieb, W.R.; Franks, N.P.
Effects of general anesthetics on the bacterial luciferase enzyme from Vibrio harveyi: an anesthetic target site with differential sensitivity
Biochemistry
29
4641-4652
1990
Vibrio harveyi
Xi, L.; Cho, K.W.; Herndon, M.E.; Tu, S.C.
Elicitation of an oxidase activity in bacterial luciferase by site-directed mutation of a noncatalytic residue
J. Biol. Chem.
265
4200-4203
1990
Vibrio harveyi
Escher, A.; O'Kane, D.J.; Lee, J.; Szalay, A.A.
Bacterial luciferase alpha beta fusion protein is fully active as a monomer and highly sensitive in vivo to elevated temperature
Proc. Natl. Acad. Sci. USA
86
6528-6532
1989
Vibrio harveyi
Angell, P.; Langley, D.; Chamberlain, A.H.L.
Localization of luciferase in luminous marine bacteria by gold immunocytochemical labelling
FEMS Microbiol. Lett.
65
177-182
1989
Aliivibrio fischeri, Vibrio harveyi
-
Kirchner, G.; Roberts, J.L.; Gustafson, G.D.; Ingolia, T.D.
Active bacterial luciferase from a fused gene: expression of a Vibrio harveyi luxAB translational fusion in bacteria, yeast and plant cells
Gene
81
349-354
1989
Vibrio harveyi
Kurfurst, M.; Macheroux, P.; Ghisla, S.; Hastings, J.W.
Bioluminescence emission of bacterial luciferase with 1-deaza-FMN. Evidence for the noninvolvement of N(1)-protonated flavin species as emitters
Eur. J. Biochem.
181
453-457
1989
Vibrio harveyi
Colepicolo, P.; Nicolas, M.T.; Bassot, J.M.; Hastings, J.W.
Expression and localization of bacterial luciferase determined by imunogold labeling
Arch. Microbiol.
152
72-76
1989
Vibrio harveyi
-
Peabody, D.S.; Andrews, C.L.; Escudero, K.W.; Devine, J.H.; Baldwin, T.O.; Bear, D.G.
A plasmid vector and quantitative techniques for the study of transcription termination in Escherichia coli using bacterial luciferase
Gene
75
289-296
1989
Vibrio harveyi
Miyamoto, C.M.; Boylan, M.; Graham, A.F.; Meighen, E.A.
Organization of the lux structural genes of Vibrio harveyi. Expression under the T7 bacteriophage promoter, mRNA analysis, and nucleotide sequence of the luxD gene
J. Biol. Chem.
263
13393-13399
1988
Vibrio harveyi
Karp, M.
Expression of bacterial luciferase genes from Vibrio harveyi in Bacillus subtilis and in Escherichia coli
Biochim. Biophys. Acta
1007
84-90
1989
Vibrio harveyi
Sugihara, J.; Baldwin, T.O.
Effects of 3 end deletions from the Vibrio harveyi luxB gene on luciferase subunit folding and enzyme assembly: generation of temperature-sensitive polypeptide folding mutants
Biochemistry
27
2872-2880
1988
Vibrio harveyi
Waddle, J.J.; Johnston, T.C.; Baldwin, T.O.
Polypeptide folding and dimerization in bacterial luciferase occur by a concerted mechanism in vivo
Biochemistry
26
4917-4921
1987
Vibrio harveyi
Baldwin, T.O.; Holzman, T.F.; Holzman, R.B.; Riddle, V.A.
Purification of bacterial luciferase by affinity methods
Methods Enzymol.
133
98-108
1986
Aliivibrio fischeri, Photobacterium phosphoreum, Vibrio harveyi
Koncz, C.; Olsson, O.; Langridge, W.H.R.; Schell, J.; Szalay, A.A.
Expression and assembly of functional bacteria luciferase in plants
Proc. Natl. Acad. Sci. USA
84
131-135
1987
Vibrio harveyi
Vervoort, J.; Muller, F.; Lee, J.; van den Berg, W.A.M.; Moonen, C.T.W.
Identificartions of the true carbon-13 nuclear magnetic resonance spectrum of the stable intermediate II in bacterial luciferase
Biochemistry
25
8062-8067
1986
Vibrio harveyi
-
Johnston, T.C.; Thompson, R.B.; Baldwin, T.O.
Nucleotide sequence of the luxB gene of Vibrio harveyi and the complete amino acid sequence of the beta subunit of bacterial luciferase
J. Biol. Chem.
261
4805-4811
1986
Vibrio harveyi
Cohn, D.H.; Milcham, A.J.; Simon, M.I.; Nealson, K.H.; Rausch, S.K.; Bonam, D.; Baldwin, T.O.
Nucleotide sequence of the luxA gene of Vibrio harveyi and the complete amino acid sequence of the alpha subunit of bacterial luciferase
J. Biol. Chem.
260
6139-6146
1985
Vibrio harveyi
Gupta, S.C.; O'Brien, D.; Hastings, J.W.
Expression of the cloned subunits of bacterial luciferase from separate replicons
Biochem. Biophys. Res. Commun.
127
1007-1011
1985
Vibrio harveyi
Swanson, R.; Weaver, L.H.; Remington, S.J.; Matthews, B.W.; Baldwin, T.O.
Crystals of luciferase from Vibrio harveyi. A preliminary characterization
J. Biol. Chem.
260
1287-1289
1985
Vibrio harveyi
Fried, A.; Tu, S.C.
Affinity labeling of the aldehyde site of bacterial luciferase
J. Biol. Chem.
259
10754-10759
1984
Vibrio harveyi
Baldwin, T.O.; Berends, T.; Bunch, T.A.; Holzman, T.F.; Rausch, S.K.; Shamansky, L.; Treat, M.L.; Ziegler, M.M.
Cloning of the luciferase structural genes from Vibrio harveyi and expression of bioluminescence in Escherichia coli
Biochemistry
23
3663-3667
1984
Vibrio harveyi
Holzman, T.F.; Baldwin, T.O.
Reversible Inhibition of the bacterial luciferase catalyzed bioluminescence reaction by aldehyde substrate: Kinetic mechanism and Ligand effects
Biochemistry
22
2838-2846
1983
Vibrio harveyi
-
Wienhausen, G.K.; Kricka, L.J.; Hinkley, J.E.; DeLuca, M.
Properties of bacterial luciferase/NADH:FMN oxidoreductase and firefly luciferase immobilized onto sepharose
Appl. Biochem. Biotechnol.
7
463-473
1982
Vibrio harveyi
Cohn, D.H.; Ogden, R.C.; Abelson, J.N.; Baldwin, T.O.; Nealson, K.H.; Simon, M.I.; Mileham, A.J.
Cloning of the Vibrio harveyi luciferase genes: use of a synthetic oligonucleotide probe
Proc. Natl. Acad. Sci. USA
80
120-123
1983
Vibrio harveyi
Tu, S.C.
Isolation and properties of bacterial luciferase intermediates containing different oxygenated flavins
J. Biol. Chem.
257
3719-3725
1982
Vibrio harveyi
Kurfurst, M.; Ghisla, S.; Presswood, R.; Hastings, J.W.
Structure and catalytic inactivity of the bacterial luciferase neutral flavin radical
Eur. J. Biochem.
123
355-361
1982
Vibrio harveyi
Welches, W.R.; Baldwin, T.O.
Active center studies on bacterial luciferase: modification of the enzyme with 2,4-dinitrofluorobenzene
Biochemistry
20
512-517
1981
Vibrio harveyi
Holzman, T.F.; Riley, P.L.; Baldwin, T.O.
Inactivation of luciferase from the Luminous marine bacterium Beneckea harveyi by proteases: evidence for a protease labile region and properties of the protein following inactivation
Arch. Biochem. Biophys.
205
554-563
1980
Vibrio harveyi
Hastings, J.W.; Presswood, R.P.
Bacterial luciferase: FMNH2-aldehyde oxidase
Methods Enzymol.
53
558-570
1978
Vibrio harveyi
Baumstark, A.L.; Cline, T.W.; Hastings, J.W.
Reversible steps in the reaction of aldehydes with bacterial luciferase intermediates
Arch. Biochem. Biophys.
193
449-455
1979
Vibrio harveyi
Hastings, J.W.; Baldwin, T.O.; Nicoli, M.Z.
Bacterial luciferase: Assay, purification, and properties
Methods Enzymol.
57
135-152
1978
Aliivibrio fischeri, Photobacterium phosphoreum, Vibrio harveyi
-
Tu, S.C.
Preparation of the subunits of bacterial luciferase
Methods Enzymol.
57
171-174
1978
Vibrio harveyi
-
Meighen.E.A.
Preparation of luciferases containing chemically modified subunits
Methods Enzymol.
57
174-181
1978
Vibrio harveyi
-
Hastings, J.W.
Bacterial bioluminescence light emission in the mixed function oxidation of reduced flavin and fatty aldehyde
CRC Crit. Rev. Biochem.
5
163-184
1978
Aliivibrio fischeri, Photobacterium leiognathi, Vibrio harveyi
Becvar, J.E.; Tu, S.C.; Hastings, J.W.
Activity and stability of the luciferase-flavin intermediate
Biochemistry
17
1807-1812
1978
Aliivibrio fischeri, Vibrio harveyi
Tu, S.C.; Wu, C.W.; Hastings, J.W.
Structural studies on bacterial luciferase using energy transfer and emission anisotropy
Biochemistry
17
987-993
1978
Vibrio harveyi
Mangold, A.; Langerman, N.
The enthalpy of oxidation of flavin mononucleotide. Temperature dependence of in vitro bacterial luciferase bioluminescence
Arch. Biochem. Biophys.
169
126-133
1975
Vibrio harveyi
Noland, B.W.; Lawrence, J.D.; Baldwin, T.O.
Folding, stability, and physical properties of the alpha subunit of bacterial luciferase
Biochemistry
38
16136-16145
1999
Vibrio harveyi
Choi, H.; Tang, C.K.; Tu, S.C.
Catalytically active forms of the individueal subunits of Vibrio harveyi luciferase and their kinetic properties
J. Biol. Chem.
270
16813-16819
1995
Vibrio harveyi
Huang, S.; Tu, S.C.
Identification and Characterization of a catalytic base in bacterial luciferase by chemical rescue of a dark mutant
Biochemistry
36
14609-14615
1997
Vibrio harveyi
Francisco, W.A.; Abu-Soud, H.M.; Baldwin, T.O.; Raushel, F.M.
Interaction of bacterial luciferase with aldehyde substrates and inhibitors
J. Biol. Chem.
268
24734-24741
1993
Vibrio harveyi
Shanker, R.; Atkins, W.M.
Luciferase-dependent, cytochrome P-450-catalyzed dehalogenation in genetically engeneered Pseudomonas
Biotechnol. Prog.
12
474-479
1996
Vibrio harveyi
Sinclair, J.F.; Waddle, J.J.; Waddill, E.F.; Baldwin, T.O.
Purified native subunits of bacterial luciferase are active in the bioluminescence reaction but fail to assemble into the alphabeta structure
Biochemistry
32
5036-5044
1993
Vibrio harveyi
Moore, C.; Lei, B.; Tu, S.C.
Relationship between the conserved alpha subunit arginine 107 and effects of phosphate on the activity and stability of Vibrio harveyi luciferase
Arch. Biochem. Biophys.
370
45-50
1999
Vibrio harveyi
Sparks, J.M.; Baldwin, T.O.
Functional implications of the unstructered loop in the (beta/alpha)8 barrel structure of the bacterial luciferase alpha subunit
Biochemistry
40
15436-15443
2001
Vibrio harveyi
Fisher, A.J.; Thompson, T.P.; Baldwin, T.O.; Rayment, I.
The 1.5-A resolution crystal structure of bacterial luciferase in low salt conditions
J. Biol. Chem.
271
21956-29168
1996
Vibrio harveyi
Szittner, R.; Jansen, G.; Thomas, D.Y.; Meighen, E.
Bright stable luminescent yeast using bacterial luciferase as a sensor
Biochem. Biophys. Res. Commun.
309
66-70
2003
Vibrio harveyi
Low, J.C.; Tu, S.C.
Functional roles of conserved residues in the unstructured loop of Vibrio harveyi bacterial luciferase
Biochemistry
41
1724-1731
2002
Vibrio harveyi
Inlow, J.K.; Baldwin, T.O.
Mutational analysis of the subunit interface of Vibrio harveyi bacterial luciferase
Biochemistry
41
3906-3915
2002
Vibrio harveyi
Lin, L.Y.C.; Sulea, T.; Szittner, R.; Kor, C.; Purisima, E.O.; Meighen, E.A.
Implications of the reactive thiol and the proximal non-proline cis-peptide bond in the structure and function of Vibrio harveyi luciferase
Biochemistry
41
9938-9945
2002
Vibrio harveyi
Jeffers, C.E.; Nichols, J.C.; Tu, S.C.
Complex formation between Vibrio harveyi luciferase and monomeric NADPH:FMN oxidoreductase
Biochemistry
42
529-534
2003
Vibrio harveyi
Lei, B.; Ding, Q.; Tu, S.C.
Identity of the emitter in the bacterial luciferase luminescence reaction: binding and fluorescence quantum yield studies of 5-decyl-4a-hydroxy-4a,5-dihydroriboflavin-5'-phosphate as a model
Biochemistry
43
15975-15982
2004
Vibrio harveyi
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
Biochemistry
43
3183-3194
2004
Vibrio harveyi
Szpilewska, H.; Czyz, A.; Wegrzyn, G.
Experimental evidence for the physiological role of bacterial luciferase in the protection of cells against oxidative stress
Curr. Microbiol.
47
379-382
2003
Vibrio harveyi
Roberts, E.A.; Clark, A.; Friedman, R.L.
Bacterial luciferase is naturally destabilized in Mycobacterium tuberculosis and can be used to monitor changes in gene expression
FEMS Microbiol. Lett.
243
243-249
2005
Vibrio harveyi
Li, X.; Tu, S.C.
Activity coupling of Vibrio harveyi luciferase and flavin reductase (FRP): oxygen as a probe
Arch. Biochem. Biophys.
454
26-31
2006
Vibrio harveyi
Li, C.H.; Tu, S.C.
Active site hydrophobicity is critical to the bioluminescence activity of Vibrio harveyi luciferase
Biochemistry
44
12970-12977
2005
Vibrio harveyi
Li, C.H.; Tu, S.C.
Probing the functionalities of alphaGlu328 and alphaAla74 of Vibrio harveyi luciferase by site-directed mutagenesis and chemical rescue
Biochemistry
44
13866-13873
2005
Vibrio harveyi
Huang, S.; Tu, S.
Effects of iodide on the fluorescence and activity of the hydroperoxyflavin intermediate of Vibrio harveyi luciferase
Photochem. Photobiol.
81
425-430
2005
Vibrio harveyi
Tu, S.C.
Activity coupling and complex formation between bacterial luciferase and flavin reductases
Photochem. Photobiol. Sci.
7
183-188
2008
Aliivibrio fischeri, Vibrio harveyi
Campbell, Z.T.; Baldwin, T.O.
Fre Is the Major Flavin Reductase Supporting Bioluminescence from Vibrio harveyi Luciferase in Escherichia coli
J. Biol. Chem.
284
8322-8328
2009
Vibrio harveyi
Campbell, Z.T.; Weichsel, A.; Montfort, W.R.; Baldwin, T.O.
Crystal structure of the bacterial luciferase/flavin complex provides insight into the function of the beta subunit
Biochemistry
48
6085-6094
2009
Vibrio harveyi (P07740)
Campbell, Z.T.; Baldwin, T.O.
Two lysine residues in the bacterial luciferase mobile loop stabilize reaction intermediates
J. Biol. Chem.
284
32827-32834
2009
Vibrio harveyi
Waidmann, M.; Bleichrodt, F.; Laslo, T.; Riedel, C.
Bacterial luciferase reporters: the swiss army knife of molecular biology
Bioeng. Bugs
2
8-16
2011
Aliivibrio fischeri, Photobacterium leiognathi, Photobacterium phosphoreum, Vibrio harveyi, Photorhabdus laumondii subsp. laumondii, Photorhabdus laumondii subsp. laumondii TT01, Photobacterium phosphoreum ATCC 11040, Photobacterium leiognathi ATCC 25521, Vibrio harveyi ATCC BAA1116
Campbell, Z.T.; Baldwin, T.O.; Miyashita, O.
Analysis of the bacterial luciferase mobile loop by replica-exchange molecular dynamics
Biophys. J.
99
4012-4019
2010
Vibrio harveyi
Close, D.M.; Patterson, S.S.; Ripp, S.; Baek, S.J.; Sanseverino, J.; Sayler, G.S.
Autonomous bioluminescent expression of the bacterial luciferase gene cassette (lux) in a mammalian cell line
PLoS ONE
5
e12441
2010
Vibrio harveyi
Melkina, O.; Goryanin, I.; Manukhov, I.; Zavilgelskii, G.
Trigger factor-dependent refolding of bacterial luciferases in Escherichia coli: Kinetics, efficiency, and effect of bichaperone system
Mol. Biol.
47
435-439
2013
Aliivibrio fischeri, Photobacterium leiognathi, Photorhabdus luminescens, Vibrio harveyi
-
Ke, D.; Tu, S.C.
Activities, kinetics and emission spectra of bacterial luciferase-fluorescent protein fusion enzymes
Photochem. Photobiol.
87
1346-1353
2011
Vibrio harveyi
Kim, T.; Spiegel, D.
Serendipitous discovery of two highly selective inhibitors of bacterial luciferase
Tetrahedron
69
7692-7698
2013
Vibrio harveyi, Vibrio harveyi BB120
-
Born, Y.; Fieseler, L.; Thoeny, V.; Leimer, N.; Duffy, B.; Loessner, M.J.
Engineering of bacteriophages Y2-dpoL1-C and Y2-luxAB for efficient control and rapid detection of the fire blight pathogen, Erwinia amylovora
Appl. Environ. Microbiol.
83
e00341
2017
Vibrio harveyi
Melkina, O.E.; Goryanin, I.I.; Manukhov, I.V.; Baranova, A.V.; Kolb, V.A.; Svetlov, M.S.; Zavilgelsky, G.B.
Trigger factor assists the refolding of heterodimeric but not monomeric luciferases
Biochemistry (Moscow)
79
62-68
2014
Photobacterium leiognathi, Photobacterium leiognathi (P07740 and P07739), Vibrio harveyi (P07740 and P07739), Aliivibrio fischeri (P19907 and P19908), Aliivibrio fischeri MJ-1 (P19907 and P19908)
Brodl, E.; Ivkovic, J.; Tabib, C.R.; Breinbauer, R.; Macheroux, P.
Synthesis of alpha,beta-unsaturated aldehydes as potential substrates for bacterial luciferases
Bioorg. Med. Chem.
25
1487-1495
2017
Aliivibrio fischeri, Photobacterium leiognathi, Vibrio harveyi, Vibrio harveyi ATCC 14126, Aliivibrio fischeri ATCC 7744
Jiang, T.; Wang, W.; Wu, X.; Wu, W.; Bai, H.; Ma, Z.; Shen, Y.; Yang, K.; Li, M.
Discovery of new substrates for LuxAB bacterial bioluminescence
Chem. Biol. Drug Des.
88
197-208
2016
Vibrio harveyi
Tinikul, R.; Lawan, N.; Akeratchatapan, N.; Pimviriyakul, P.; Chinantuya, W.; Suadee, C.; Sucharitakul, J.; Chenprakhon, P.; Ballou, D.P.; Entsch, B.; Chaiyen, P.
Protonation status and control mechanism of flavin-oxygen intermediates in the reaction of bacterial luciferase
FEBS J.
288
3246-3260
2021
Vibrio harveyi (P07740 and P07739)
Nemtseva, E.V.; Gulnov, D.V.; Gerasimova, M.A.; Sukovatyi, L.A.; Burakova, L.P.; Karuzina, N.E.; Melnik, B.S.; Kratasyuk, V.A.
Bacterial luciferases from Vibrio harveyi and Photobacterium leiognathi demonstrate different conformational stability as detected by time-resolved fluorescence spectroscopy
Int. J. Mol. Sci.
22
14994
2021
Vibrio harveyi (P07740 and P07739), Photobacterium leiognathi (P09140 and P09141)
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