Information on EC 1.13.12.4 - lactate 2-monooxygenase

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The expected taxonomic range for this enzyme is: Mycobacterium

EC NUMBER
COMMENTARY
1.13.12.4
-
RECOMMENDED NAME
GeneOntology No.
lactate 2-monooxygenase
REACTION
REACTION DIAGRAM
COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
(S)-lactate + O2 = acetate + CO2 + H2O
show the reaction diagram
-
-
-
-
REACTION TYPE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
oxidation
-
-
-
-
oxidative decarboxylation
-
-
-
-
redox reaction
-
-
-
-
reduction
-
-
-
-
PATHWAY
KEGG Link
MetaCyc Link
Pyruvate metabolism
-
SYSTEMATIC NAME
IUBMB Comments
(S)-lactate:oxygen 2-oxidoreductase (decarboxylating)
A flavoprotein (FMN).
SYNONYMS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
EC 1.1.3.2
-
-
formerly
-
L-lactate monooxygenase
-
-
-
-
L-lactate-2-monooxygenase
-
-
-
-
lactate monooxygenase
-
-
-
-
lactate oxidase
-
-
-
-
lactate oxidative decarboxylase
-
-
-
-
lactate oxygenase
-
-
-
-
lactic oxidase
-
-
-
-
lactic oxygenase
-
-
-
-
CAS REGISTRY NUMBER
COMMENTARY
9028-72-2
-
ORGANISM
COMMENTARY
LITERATURE
SEQUENCE CODE
SEQUENCE DB
SOURCE
Gram-negative soil bacterium
strain KY6
-
-
Manually annotated by BRENDA team
Gram-negative soil bacterium KY6
strain KY6
-
-
Manually annotated by BRENDA team
strain ATCC 14468
-
-
Manually annotated by BRENDA team
SUBSTRATE
PRODUCT                      
REACTION DIAGRAM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
(Substrate)
LITERATURE
(Substrate)
COMMENTARY
(Product)
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
(R)-lactate + 2,6-dichlorophenolindophenol
?
show the reaction diagram
Gram-negative soil bacterium, Gram-negative soil bacterium KY6
-
no decarboxylation
-
-
?
(R)-lactate + O2
pyruvate + H2O2
show the reaction diagram
-
apoenzyme with 6,7-dimethyl FMN
-
-
?
(S)-lactate + 2,6-dichlorophenolindophenol
pyruvate + ?
show the reaction diagram
-
no decarboxylation
-
-
?
(S)-lactate + 2,6-dichlorophenolindophenol
pyruvate + ?
show the reaction diagram
Gram-negative soil bacterium, Gram-negative soil bacterium KY6
-
no decarboxylation
-
-
?
(S)-lactate + methylene blue
pyruvate + ?
show the reaction diagram
-
-
-
-
?
(S)-lactate + O2
acetate + CO2 + H2O
show the reaction diagram
-
-
-
?
(S)-lactate + O2
acetate + CO2 + H2O
show the reaction diagram
-
-
-
?
(S)-lactate + O2
acetate + CO2 + H2O
show the reaction diagram
-
-
-
?
(S)-lactate + O2
acetate + CO2 + H2O
show the reaction diagram
-
-
-
?
(S)-lactate + O2
acetate + CO2 + H2O
show the reaction diagram
-
-
-
?
(S)-lactate + O2
acetate + CO2 + H2O
show the reaction diagram
-
-
-
?
(S)-lactate + O2
acetate + CO2 + H2O
show the reaction diagram
-
-
-
?
(S)-lactate + O2
acetate + CO2 + H2O
show the reaction diagram
-
-
-
?
(S)-lactate + O2
acetate + CO2 + H2O
show the reaction diagram
-
-
-
?
(S)-lactate + O2
acetate + CO2 + H2O
show the reaction diagram
-
-
-
?
(S)-lactate + O2
acetate + CO2 + H2O
show the reaction diagram
-
-
-
?
(S)-lactate + O2
acetate + CO2 + H2O
show the reaction diagram
-
-
-
?
(S)-lactate + O2
acetate + CO2 + H2O
show the reaction diagram
-
-
-
-
?
(S)-lactate + O2
pyruvate + H2O2
show the reaction diagram
-
apoenzyme with 6,7-dimethyl FMN
-
-
?
(S)-lactate + O2
pyruvate + H2O2
show the reaction diagram
-
G99A-mutant, no decarboxylation
-
-
?
(S)-malate + O2
oxaloacetate + H2O2
show the reaction diagram
-
-
-
?
2-hydroxy-3-butynoate + O2
?
show the reaction diagram
-
-
-
-
?
2-hydroxy-3-methylvalerate + O2
?
show the reaction diagram
-
-
-
-
?
2-hydroxybutyrate + O2
?
show the reaction diagram
-
-
-
-
?
2-hydroxycaproate + O2
?
show the reaction diagram
-
-
-
-
?
2-hydroxyisovalerate + O2
?
show the reaction diagram
-
-
-
-
?
2-hydroxyvalerate + O2
?
show the reaction diagram
-
-
-
-
?
beta-chlorolactate + electron acceptor
pyruvate + Cl- + H2O
show the reaction diagram
-
under anaerobic conditions
-
?
beta-chlorolactate + O2
chloroacetate + CO2 + H2O
show the reaction diagram
-
-
-
?
beta-phenyllactate + O2
?
show the reaction diagram
-
-
-
-
?
beta-phenyllactate + O2
?
show the reaction diagram
-
-
-
-
?
DL-2-hydroxy-3-butynoate + O2
?
show the reaction diagram
-
-
-
-
?
DL-mandelate + O2
?
show the reaction diagram
-
-
-
-
?
glycolate + 2,6-dichlorophenolindophenol
?
show the reaction diagram
Gram-negative soil bacterium, Gram-negative soil bacterium KY6
-
no decarboxylation
-
-
?
glycolate + electron acceptor
glyoxylate + electron donor
show the reaction diagram
-
under anaerobic conditions, leads to two different adducts depending on the initial abstraction of the re- or si-alpha-hydrogen
electron donor is H2O2 under aerobic conditions
?
glyoxylate + O2
oxalate + H2O2
show the reaction diagram
-
-
-
?
L-2-hydroxyisocaproate + 2,6-dichlorophenolindophenol
?
show the reaction diagram
Gram-negative soil bacterium, Gram-negative soil bacterium KY6
-
no decarboxylation
-
-
?
L-lactate + O2
acetate + CO2 + H2O
show the reaction diagram
-
-
-
-
?
L-malic acid + 2,6-dichlorophenolindophenol
?
show the reaction diagram
Gram-negative soil bacterium
-
no decarboxylation
-
-
?
lactate + 2,6-dichlorophenolindophenol
?
show the reaction diagram
Gram-negative soil bacterium, Gram-negative soil bacterium KY6
-
no decarboxylation
-
-
?
NATURAL SUBSTRATES
NATURAL PRODUCTS
REACTION DIAGRAM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
(Substrate)
LITERATURE
(Substrate)
COMMENTARY
(Product)
LITERATURE
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
(S)-lactate + O2
acetate + CO2 + H2O
show the reaction diagram
-
-
-
?
(S)-lactate + O2
acetate + CO2 + H2O
show the reaction diagram
-
-
-
?
(S)-lactate + O2
acetate + CO2 + H2O
show the reaction diagram
-
-
-
?
(S)-lactate + O2
acetate + CO2 + H2O
show the reaction diagram
-
-
-
?
(S)-lactate + O2
acetate + CO2 + H2O
show the reaction diagram
-
-
-
?
(S)-lactate + O2
acetate + CO2 + H2O
show the reaction diagram
-
-
-
?
(S)-lactate + O2
acetate + CO2 + H2O
show the reaction diagram
-
-
-
?
(S)-lactate + O2
acetate + CO2 + H2O
show the reaction diagram
-
-
-
?
(S)-lactate + O2
acetate + CO2 + H2O
show the reaction diagram
-
-
-
?
(S)-lactate + O2
acetate + CO2 + H2O
show the reaction diagram
-
-
-
?
(S)-lactate + O2
acetate + CO2 + H2O
show the reaction diagram
-
-
-
?
(S)-lactate + O2
acetate + CO2 + H2O
show the reaction diagram
-
-
-
?
(S)-lactate + O2
acetate + CO2 + H2O
show the reaction diagram
-
-
-
-
?
COFACTOR
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
FMN
Gram-negative soil bacterium
-
-
FMN
-
one FMN per subunit
FMN
-
with 5-deaza FMN the enzyme undergoes reduction by (S)-lactate but not catalytic reduction by O2
FMN
-
6 mol per mol protein
FMN
-
1 mol FMN per 44000 g enzyme
riboflavin
-
24% activity of that with FMN
INHIBITORS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
(R)-lactate
Gram-negative soil bacterium
-
at concentrations higher than 10 mM
(R)-lactate
-
competitive inhibitor
(R)-lactate
-
competitive inhibitor
1-Fluoro-2,4-dinitrobenzene
-
after 2 h at 2 mM, complete inactivation by dinitrophenylation of 2 mol of histidine residues per mol of enzyme-bound FMN, competitive inhibitors such as phosphate, nitrate and alpha-hydroxymalonate decrease the rate of inactivation
2,3-Butanedione
-
90% inactivation at 50 mM after 90 min in 50 mM borate buffer, gel filtration in 50 mM phosphate buffer causes a recovery of 92% activiy
2,3-epoxypropionic acid
-
-
2-Hydroxy-3-butynoate
-
suicide substrate
2-Hydroxy-3-butynoate
-
irreversible inactivator due to a covalent modification of the bound FMN
2-Methyllactate
-
competitive inhibitor
acetate
-
competitive inhibitor
D-lactate
-
competitive
diethyl dicarbonate
-
6fold molar excess with respect to enzyme-bound FMN results in 92% inactivation after 13 min, substrate and competitive inhibitors decrease the maximum extent of inactivation to a 50%, modification of histidines
FeSO4
Gram-negative soil bacterium
-
86% inhibition at 1 mM
fluorodintrobenzene
-
complete inactivation, incorporation of 1 mol dinitrophenyl per catalytic site
formate
-
competitive inhibitor
HgCl2
Gram-negative soil bacterium
-
28% inhibition at 1 mM
iodoacetate
Gram-negative soil bacterium
-
33% inhibition at 1 mM
L-leucine
Gram-negative soil bacterium
-
29% inhibition at 1 mM
L-phenylalanine
Gram-negative soil bacterium
-
28% inhibition at 1 mM
Lactate
Gram-negative soil bacterium
-
at concentrations higher than 20 mM
MnSO4
Gram-negative soil bacterium
-
47% inhibition at 1 mM
oxalate
-
transition state analog of a carbanion form of the substrate
oxalate
-
inhibition is reversible in the absence and irreversible in the presence of light
Oxamate
-
competitive inhibitor
p-chloromercuribenzoate
Gram-negative soil bacterium
-
21% inhibition at 1 mM
p-chloromercuribenzoate
-
100% inhibition at 1.1 mM at pH 8 and 25C after 30 min, excess of 2-mercaptoethanol protects
Phenylglyoxal
-
2 equivalents per subunit are required for total inactivation, in presence of competitive inhibitors inactivation is markedly reduced
phosphate
-
competitive inhibitor
Tetranitromethane
-
20 min at pH 8.0 and 1.5 mM; rapid and irreversible inactivation at 30C, nitration of a single tyrosine per subunit, competitive inhibitors such as acetate, (R)-lactate or oxalate protect from inactivation
ZnSO4
Gram-negative soil bacterium
-
28% inhibition at 1 mM
ACTIVATING COMPOUND
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
EDTA
Gram-negative soil bacterium
-
40% activation at 1 mM
KM VALUE [mM]
KM VALUE [mM] Maximum
SUBSTRATE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
0.14
-
(R)-lactate
Gram-negative soil bacterium
-
-
0.014
-
(S)-Lactate
-
calculated constant, G99A mutant
0.02
-
(S)-Lactate
-
observed constant, G99A mutant
0.44
-
(S)-Lactate
-
Y44F mutant
6
-
(S)-Lactate
-
Y152F mutant
10
-
(S)-Lactate
-
R293K mutant
20
-
(S)-Lactate
-
H290Q mutant
22
-
(S)-Lactate
-
wild-type
22.3
-
(S)-Lactate
-
-
34
-
(S)-Lactate
-
-
50
-
beta-phenyl-(S)-lactate
-
-
1.41
-
DL-2-hydroxy-3-butynoate
-
-
2
-
DL-2-hydroxy-3-butynoate
-
-
25
-
L-2-hydroxyisovalerate
-
-
11.3
-
L-alpha-hydroxy-beta-methylvalerate
-
-
0.2
-
Lactate
Gram-negative soil bacterium
-
-
0.055
-
O2
-
calculated constant, wild-type
0.071
-
O2
-
observed constant, wild-type
0.071
-
O2
-
wild-type
0.83
-
O2
-
observed constant, G99A mutant
0.92
-
O2
-
calculated constant, G99A mutant
TURNOVER NUMBER [1/s]
TURNOVER NUMBER MAXIMUM[1/s]
SUBSTRATE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
0.0148
-
(S)-Lactate
-
-
Ki VALUE [mM]
Ki VALUE [mM] Maximum
INHIBITOR
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
4
-
(R)-lactate
-
-
23
-
2,3-Butanedione
-
-
6.4
-
2,3-epoxypropionic acid
-
-
4
-
2-methyl-lactate
-
-
0.4
-
nitrate
-
-
-
0.01
-
oxalate
-
-
SPECIFIC ACTIVITY [µmol/min/mg]
SPECIFIC ACTIVITY MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
pH OPTIMUM
pH MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
7.7
-
Gram-negative soil bacterium
-
-
TEMPERATURE OPTIMUM
TEMPERATURE OPTIMUM MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
70
-
Gram-negative soil bacterium
-
-
MOLECULAR WEIGHT
MOLECULAR WEIGHT MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
204000
-
Gram-negative soil bacterium
-
gel filtration
300000
-
-
sucrose density gradient centrifugation
340000
-
-
gel filtration
341000
-
-
sedimentation velocity data
345000
-
-
sedimentation-equilibrium analysis
352000
-
-
low speed sedimentation
370000
-
-
sedimentation equilibrium analysis
SUBUNITS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
?
Gram-negative soil bacterium
-
? * 45000, SDS-PAGE
?
-
? * 43072, including FMN, amino acid analysis
?
Gram-negative soil bacterium KY6
-
? * 45000, SDS-PAGE
-
hexamer
-
6 * 56000, flavin and terminal amino acid analysis; 6 * 58600, electron microscopic analysis
hexamer
-
6 * 54000-57000, sedimentation velocity analysis and diffusion coefficient determination
octamer
-
8 * 43000, amino acid analysis
octamer
-
8 * 43655, amino acid analysis, per mol of FMN
Crystallization/COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
pH STABILITY
pH STABILITY MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
5.7
9.5
Gram-negative soil bacterium
-
stable for 24 h at 4C
TEMPERATURE STABILITY
TEMPERATURE STABILITY MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
0
45
Gram-negative soil bacterium
-
stable for 10 min
55
-
Gram-negative soil bacterium
-
50% activity after 10 min
55
-
-
20 min stable
GENERAL STABILITY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
dissociation of subunits does not occur in the presence of 8 M urea and 100 mM 2-mercaptoethanol
-
dissociation of subunits occurs in the presence of 5 M guanidine hydrochloride and 100 mM 2-mercaptoethanol
-
STORAGE STABILITY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
4C, 80% saturated ammonium sulfate in the dark
-
4C, crystalline suspension in 1 M sodium acetate buffer, pH 5.4, in the dark
-
5C, crystalline suspension in ammonium sulfate, several months, no loss of activity
-
0C, crystalline suspension at 2-10 mg per ml for at least 6 months without loss of activity in the dark
-
4C, 1 M sodium acetate buffer, pH 5.7, in the dark
-
Purification/COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
-
Gram-negative soil bacterium
-
Cloned/COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
expressed in Escherichia coli
-
expressed in Escherichia coli BL21
-
mutant enzyme H290Q expressed in Escherichia coli
-
ENGINEERING
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
G99A
-
(S)-lactate binds tighter to the G99A-mutant than to wild-type
H290Q
-
the ability of L-lactate to reduce the enzyme flavin is abolished, whereas reoxidation of reduced enzyme with oxygen proceeds at a rate like that found in the wild type enzyme. Unlike the situation with wild type enzyme, where the transition state analog oxalate is bound tightly in a two-step reaction involving proton uptake from solution, the mutant enzyme binds oxalate weakly, in a single step reaction. Replacing the histidine has a significant effect on the ability of the enzyme to stabilize the flavin N(5)-sulfite adduct. Sulfite is bound at least 1000fold weaker than it is in the wild type enzyme
K266M
-
the rate of reduction with (S)-lactate is decreased compared with that of the wild-type enzyme, the mutant enzyme is virtually inactive
R293K
-
uncoupled reaction, no decarboxylation
Y152F
-
nearly as active as wild-type
Y44F
-
uncoupled reaction, no decarboxylation