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Information on EC 1.14.14.3 - bacterial luciferase and Organism(s) Photorhabdus luminescens and UniProt Accession P19840
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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: Photorhabdus luminescens
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
LuxA
Vibrio fischeri luciferase
-
-
-
-
bacterial luciferase
-
-
-
-
luciferase
-
-
-
-
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
decanal + FMNH + O2
decanoic acid + FMN + H2O + light
-
-
-
-
ir
dodecanal + FMNH + O2
dodecanoic acid + FMN + H2O + light
-
-
-
-
ir
myristic aldehyde + FMNH + O2
myristic acid + FMN + H2O + light
-
-
-
-
?
n-decanal + FMNH2 + O2
n-decanoate + FMN + H2O + hn
-
3-step process via H2O2 as intermediate
generation of blue-green light of wavelength 490 nm
-
ir
octanal + FMNH + O2
octanoic acid + FMN + H2O + light
-
-
-
-
ir
decanal + FMNH2 + O2
decanoic acid + FMN + H2O + hv
-
-
-
?
decanal + FMNH2 + O2
decanoic acid + FMN + H2O + hv
-
-
light emission at 490 nm
-
?
RCHO + FMNH2 + O2
RCOOH + FMN + H2O + hn
-
3-step process via H2O2 as intermediate
generation of blue-green light of wavelength 490 nm
-
ir
RCHO + FMNH2 + O2
RCOOH + FMN + H2O + hn
-
long-chain aldehydes
long-chain fatty acids, bioluminescence reaction
-
ir
additional information
?
-
-
the decay rate of the enzyme is determined by residue Glu175 of the central region of the LuxA subunit, distinction between slow and fast decay luciferases is primarily due to differences in aldehyde affinity and in the decomposition of the luciferase-flavin-oxygen intermediate
-
-
?
additional information
?
-
-
substrate specificity and quantum yield of mutant E175G as a function of aldehyde chain length
-
-
?
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
decanal + FMNH + O2
decanoic acid + FMN + H2O + light
-
-
-
-
ir
additional information
?
-
-
the decay rate of the enzyme is determined by residue Glu175 of the central region of the LuxA subunit, distinction between slow and fast decay luciferases is primarily due to differences in aldehyde affinity and in the decomposition of the luciferase-flavin-oxygen intermediate
-
-
?
RCHO + FMNH2 + O2
RCOOH + FMN + H2O + hn
-
3-step process via H2O2 as intermediate
generation of blue-green light of wavelength 490 nm
-
ir
RCHO + FMNH2 + O2
RCOOH + FMN + H2O + hn
-
long-chain aldehydes
long-chain fatty acids, bioluminescence reaction
-
ir
COFACTOR
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
additional information
-
the enzyme shows enhanced affinity for chaperone ClpA
-
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
additional information
additional information
-
tetradecanal + FMNH + O2 (possibly) turnover rate determined by measuring the half-time for decay of luminescence between 70% and 35% of the maximum intensity, values depending on chain-length of aldehyde
-
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
ORGANISM
COMMENTARY
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
physiological function
physiological function
-
expression in Staphylococcus aureus Xen29. In the absence of antibiotics, staphylococcal bioluminescence increases over time until a maximum after ca. 6 h of growth, and subsequently decreases to the detection threshold after 24 h of growth. Up to minimal inhibitory concentrations of the antibiotics vancomycin, ciprofloxacin, erythromycin or chloramphenicol, bioluminescence increases according to a similar pattern up to 6 h of growth, but after 24 h, bioluminescence is higher than in the absence of antibiotics. Antibiotic pressure impacts the relation between bioluminescence per organism and bioluminescence. Under antibiotic pressure, bioluminescence is not controlled by luxA expression but by cofactors impacting the bacterial metabolic activity
physiological function
transient and stable transfection of human kidney, breast cancer, and colorectal cancer cell lines by a codon optimized lux expression cassette using viral 2A elements as linker regions. The expression product produces autobioluminescent phenotypes in all cell lines tested without the induction of cytotoxicity and allows for repeated interrogation of populations and self-directed control of bioluminescent activation with detection limits and EC50 values similar to traditional reporter systems
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). MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
SUBUNIT
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
PROTEIN VARIANTS
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
E175G
-
random mutagenesis, the single point mutation leads to increased decay rate of the enzyme, 0.9% of wild-type luminescence activity
E175G/N199D
-
random mutagenesis, 0.1% of wild-type luminescence activity
TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
additional information
-
the recombinant enzyme shows reduced thermal stability in absence of chaperone Hsp100 after expression in an Escherichia coli clpA-mutant strain, lack of chaperone ClpB decreases the thermal stability of the recombinant enzyme in vivo
GENERAL STABILITY
ORGANISM
UNIPROT
LITERATURE
the wild-type enzyme belongs to the group of luciferases with slow decay, mutant E175G is turned into a luciferase with fast decay, the decay rate of the enzyme is determined by residue Glu175
-
PURIFICATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
CLONED (Commentary)
ORGANISM
UNIPROT
LITERATURE
stable expression, using a bicistronic expression vector, of wild type luxA and luxB, WTA/WTB, codon-optimized luxA and wild type luxB, COA/WTB, and codon-optimized versions of both luxA and luxB genes, COA/COB, in HEK-293 cells, expression analysis, method evaluation and optimization, highest bioluminescence by expression of both codon-optimized genes, overview
expression in Bacillus subtilis and in Escherichia coli
expression in different Escherichia coli strains, which are wild-type, or deficient in gene clpA, clpB, and clpX encoding Hsp chaperones, respectively
-
expression of wild-type and randomly generated mutants in Escherichia coli strain BL21
-
stable expression, using a bicistronic expression vector, of wild type luxA and luxB, WTA/WTB, codon-optimized luxA and wild type luxB, COA/WTB, and codon-optimized versions of both luxA and luxB genes, COA/COB, in HEK-293 cells, expression analysis, method evaluation and optimization, highest bioluminescence by expression of both codon-optimized genes, overview
RENATURED/Commentary
ORGANISM
UNIPROT
LITERATURE
slow refolding at room temperature or 35°C of thermoinactivated recominant enzyme requires the DnaK-DnaJ-GrpE-system encoding the Hsp70 chaperone, 7-8% activity after refolding over 10 min, refolding depends on the Escherichia coli strain used for recombinant expression of the luciferase, overview
-
APPLICATION
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
analysis
expression of the bacterial luciferase system in mammalian cells for generation of bioreporters for in vivo monitoring and diagnostics technology, method evaluation and optimization, overview
diagnostics
expression of the bacterial luciferase system in mammalian cells for generation of bioreporters for in vivo monitoring and diagnostics technology, method evaluation and optimization, overview
agriculture
KJ957766
optimization of fused luxAB expression, quantum yield and application as a reporter gene in plant protoplasts. Luciferase activity is dramatically increased upon use of the optimized gene and the 35S promoter compared to the original luxAB in Arabidopsis and maize cells
analysis
diagnostics
expression of the bacterial luciferase system in mammalian cells for generation of bioreporters for in vivo monitoring and diagnostics technology, method evaluation and optimization, overview
medicine
synthesis
upon expression in Bacillus subtilis cells, luciferase is substantially more thermostable than in Escherichia coli. Thermal inactivation in Bacillus subtilis at 48.5#°C behaves as a first-order reaction. In Escherichia coli, the first order rate constant of the thermal inactivation exceeds that observed in B. subtilis cells 2.9 times. In dnaK-negative strains of Bacillus subtilis, both the rates of thermal inactivation and the efficiency of refolding are similar to that observed in wild-type strains
analysis
expression of the bacterial luciferase system in mammalian cells for generation of bioreporters for in vivo monitoring and diagnostics technology, method evaluation and optimization, overview
analysis
-
high-throughput, homogeneous, bioluminescent assay for Pseudomonas aeruginosa gyrase inhibitors and other DNA-damaging agents based on a Photorhabdus luminescens luciferase operon transcriptional fusion to a promoter that responds to DNA damage caused by reduced gyrase levels and fluoroquinoline inhibition
analysis
fusion of circularly permuted Venus, a bright variant of yellow fluorescent protein, to the C-terminus of subunit LuxB to induce bioluminescence resonance energy transfer (BRET). By using decanal as added substrate, color change and ten-times enhancement of brightness is achieved in Escherichia coli upon expression. Expression of the Venus-fused luciferase in human embryonic kidney cell lines or in Nicotiana benthamiana leaves together with the substrate biosynthesis-related genes (luxC, luxD and luxE) enhances the autonomous bioluminescence
medicine
-
expression in Staphylococcus aureus Xen29. In the absence of antibiotics, staphylococcal bioluminescence increases over time until a maximum after ca. 6 h of growth, and subsequently decreases to the detection threshold after 24 h of growth. Up to minimal inhibitory concentrations of the antibiotics vancomycin, ciprofloxacin, erythromycin or chloramphenicol, bioluminescence increases according to a similar pattern up to 6 h of growth, but after 24 h, bioluminescence is higher than in the absence of antibiotics. Antibiotic pressure impacts the relation between bioluminescence per organism and bioluminescence. Under antibiotic pressure, bioluminescence is not controlled by luxA expression but by cofactors impacting the bacterial metabolic activity
medicine
transient and stable transfection of human kidney, breast cancer, and colorectal cancer cell lines by a codon optimized lux expression cassette using viral 2A elements as linker regions. The expression product produces autobioluminescent phenotypes in all cell lines tested without the induction of cytotoxicity and allows for repeated interrogation of populations and self-directed control of bioluminescent activation with detection limits and EC50 values similar to traditional reporter systems
REF.
AUTHORS
TITLE
JOURNAL
VOL.
PAGES
YEAR
ORGANISM (UNIPROT)
PUBMED ID
SOURCE
Xi, L.; Cho, K.W.; Tu, S.C.
Cloning and nucleotide sequences of lux genes and characterization of luciferase of Xenorhabdus luminescens from a human wound
J. Bacteriol.
173
1399-1405
1991
Photorhabdus luminescens
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
Li, Z.; Meighen, E.A.
The turnover of bacterial luciferase is limited by a slow decomposition of the ternary enzyme-product complex of luciferase, FMN, and fatty acid
J. Biol. Chem.
269
6640-6644
1994
Photorhabdus luminescens
Hosseinkhani, S.; Szittner, R.; Meighen, E.A.
Random mutagenesis of bacterial luciferase: critical role of Glu175 in the control of luminescence decay
Biochem. J.
385
575-580
2005
Photorhabdus luminescens
Zavilgelsky, G.B.; Kotova, V.Y.; Mazhul, M.M.; Manukhov, I.V.
Role of Hsp70 (DnaK-DnaJ-GrpE) and Hsp100 (ClpA and ClpB) chaperones in refolding and increased thermal stability of bacterial luciferases in Escherichia coli cells
Biochemistry (Moscow)
67
986-992
2002
Aliivibrio fischeri, Photorhabdus luminescens
Patterson, S.S.; Dionisi, H.M.; Gupta, R.K.; Sayler, G.S.
Codon optimization of bacterial luciferase (lux) for expression in mammalian cells
J. Ind. Microbiol. Biotechnol.
32
115-123
2005
Photorhabdus luminescens (P19839), Photorhabdus luminescens (P19840)
Moir, D.T.; Ming Di.; Opperman, T.; Schweizer, H.P.; Bowlin, T.L.
A high-throughput, homogeneous, bioluminescent assay for Pseudomonas aeruginosa gyrase inhibitors and other DNA-damaging agents
J. Biomol. Screen.
12
855-864
2007
Photorhabdus luminescens
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
-
Daghighi, S.; Sjollema, J.; Harapanahalli, A.; Dijkstra, R.J.; van der Mei, H.C.; Busscher, H.J.
Influence of antibiotic pressure on bacterial bioluminescence, with emphasis on Staphylococcus aureus
Int. J. Antimicrob. Agents
46
713-717
2015
Photorhabdus luminescens
Cui, B.; Zhang, L.; Song, Y.; Wei, J.; Li, C.; Wang, T.; Wang, Y.; Zhao, T.; Shen, X.
Engineering an enhanced, thermostable, monomeric bacterial luciferase gene as a reporter in plant protoplasts
PLoS ONE
9
e107885
2014
Photorhabdus luminescens (KJ957766), Photorhabdus luminescens
Xu, T.; Ripp, S.; Sayler, G.S.; Close, D.M.
Expression of a humanized viral 2A-mediated lux operon efficiently generates autonomous bioluminescence in human cells
PLoS ONE
9
e96347
2014
Photorhabdus luminescens (P19839 and P19840)
Gnuchikh, E.; Baranova, A.; Schukina, V.; Khaliullin, I.; Zavilgelsky, G.; Manukhov, I.
Kinetics of the thermal inactivation and the refolding of bacterial luciferases in Bacillus subtilis and in Escherichia coli differ
PLoS ONE
14
e0226576
2019
Photobacterium leiognathi (P09140 and P09141), Photobacterium leiognathi, Photorhabdus luminescens (P23146 and P19840), Photorhabdus luminescens
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