Information on EC 1.1.2.3 - L-lactate dehydrogenase (cytochrome)

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The expected taxonomic range for this enzyme is: Eukaryota, Bacteria

EC NUMBER
COMMENTARY
1.1.2.3
-
RECOMMENDED NAME
GeneOntology No.
L-lactate dehydrogenase (cytochrome)
REACTION
REACTION DIAGRAM
COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
(S)-lactate + 2 ferricytochrome c = pyruvate + 2 ferrocytochrome c + 2 H+
show the reaction diagram
-
-
-
-
(S)-lactate + 2 ferricytochrome c = pyruvate + 2 ferrocytochrome c + 2 H+
show the reaction diagram
hydride transfer mechanism
-
(S)-lactate + 2 ferricytochrome c = pyruvate + 2 ferrocytochrome c + 2 H+
show the reaction diagram
catalytic cycle, overview
-
REACTION TYPE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
oxidation
-
-
-
-
redox reaction
-
-
-
-
reduction
-
-
-
-
PATHWAY
KEGG Link
MetaCyc Link
Metabolic pathways
-
methylglyoxal degradation V
-
Pyruvate metabolism
-
SYSTEMATIC NAME
IUBMB Comments
(S)-lactate:ferricytochrome-c 2-oxidoreductase
Identical with cytochrome b2; a flavohemoprotein (FMN).
SYNONYMS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
Cyb2
Saccharomyces cerevisiae D273-10B/A1
-
-
-
cytochrome b2
-
-
-
-
cytochrome c oxido reductase
-
-
FC b2
Ogataea angusta 356, Ogataea angusta C-105, Ogataea angusta K-105
-
-
-
FC b2
Saccharomyces cerevisiae IZR-106, Saccharomyces cerevisiae IZR-42
-
-
-
flavocytochrome b
-
-
flavocytochrome b2
-
-
-
-
flavocytochrome b2
-
-
flavocytochrome b2
Ogataea angusta 356, Ogataea angusta C-105, Ogataea angusta K-105
-
-
-
flavocytochrome b2
P00175
-
flavocytochrome b2
Saccharomyces cerevisiae D273-10B/A1, Saccharomyces cerevisiae IZR-106, Saccharomyces cerevisiae IZR-42
-
-
-
flavocytochrome b2
-
-
L(+)-lactate:cytochrome c oxidoreductase
-
-
-
-
L-(+)-lactate ferricytochrome c oxidoreductase
-
-
L-lactate cytochrome c oxidoreductase
-
-
-
-
L-lactate cytochrome c oxidoreductase
-
-
L-lactate cytochrome c oxidoreductase
Ogataea angusta 356, Ogataea angusta K-105
-
-
-
L-lactate cytochrome c oxidoreductase
-
-
L-lactate cytochrome c oxidoreductase
P00175
-
L-lactate cytochrome c oxidoreductase
Saccharomyces cerevisiae IZR-106, Saccharomyces cerevisiae IZR-42
-
-
-
L-lactate cytochrome c reductase
-
-
-
-
L-lactate dehydrogenase [Cytochrome]
-
-
-
-
L-lactate ferricytochrome C oxidoreductase
-
-
-
-
L-lactate ferricytochrome C oxidoreductase
-
-
L-lactate: cytochrome c oxidoreductase
-
-
L-lactate:cytochrome c oxidoreductase
-
-
L-lactate:cytochrome c oxidoreductase
P00175
-
L-lactate:cytochrome c-oxidoreductase
-
-
L-lactate:cytochrome c-oxidoreductase
Ogataea angusta C-105
-
-
-
L-lactate:cytochrome c-oxidoreductase
-
-
L-LCR
-
-
-
-
L-LDH (FMN-dependent)
-
-
lactate dehydrogenase (cytochrome)
-
-
-
-
lactic acid dehydrogenase
-
-
-
-
lactic cytochrome c reductase
-
-
-
-
NAD+ - independent LDH
-
-
CAS REGISTRY NUMBER
COMMENTARY
9078-32-4
-
ORGANISM
COMMENTARY
LITERATURE
SEQUENCE CODE
SEQUENCE DB
SOURCE
L. Corm, before, 10 and 30 days after rooting, three isoforms
-
-
Manually annotated by BRENDA team
three isoforms
-
-
Manually annotated by BRENDA team
356; K-105
-
-
Manually annotated by BRENDA team
i.e. Pichia angusa, gene CYB2
-
-
Manually annotated by BRENDA team
Ogataea angusta 356
356
-
-
Manually annotated by BRENDA team
Ogataea angusta C-105
-
-
-
Manually annotated by BRENDA team
Ogataea angusta K-105
K-105
-
-
Manually annotated by BRENDA team
cytochrome domain expressed in Escherichia coli
-
-
Manually annotated by BRENDA team
IZR-106; IZR-42
-
-
Manually annotated by BRENDA team
Saccharomyces cerevisiae D273-10B/A1
-
-
-
Manually annotated by BRENDA team
Saccharomyces cerevisiae IZR-106
IZR-106
-
-
Manually annotated by BRENDA team
Saccharomyces cerevisiae IZR-42
IZR-42
-
-
Manually annotated by BRENDA team
flavin and heme domain expressed in Escherichia coli
-
-
Manually annotated by BRENDA team
GENERAL INFORMATION
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
metabolism
-
the pyruvate produced as a result of L-lactate oxidation is utilised by the Krebs cycle, but flavocytochrome b2 also forms part of a short respiratory electron transport chain which results in one ATP molecule being produced for every L-lactate consumed
physiological function
-
flavocytochrome b2, i.e. L-lactate cytochrome c oxidoreductase, catalyzes L-lactate oxidation at the expense of cytochrome c and enables the latter to grow on lactate as the sole carbon source
physiological function
-
the enzyme is involved in respiration and energy production
SUBSTRATE
PRODUCT                      
REACTION DIAGRAM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
(Substrate)
LITERATURE
(Substrate)
COMMENTARY
(Product)
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
(S)-2-hydroxybutyrate + ferricytochrome c
2-oxobutyrate + ferrocytochrome c
show the reaction diagram
-
-
-
-
?
(S)-2-hydroxyhexanoate + ferricytochrome c
2-oxohexanoate + ferrocytochrome c
show the reaction diagram
-
-
-
-
?
(S)-2-hydroxyoctanoate + ferricytochrome c
2-oxooctanoate + ferrocytochrome c
show the reaction diagram
-
-
-
-
?
(S)-2-hydroxyvalerate + ferricytochrome c
2-oxovalerate + ferrocytochrome c
show the reaction diagram
-
-
-
-
?
(S)-lactate + 2 ferricytochrome c
pyruvate + 2 ferrocytochrome c + 2 H+
show the reaction diagram
-
-
-
-
(S)-lactate + 2 ferricytochrome c
pyruvate + 2 ferrocytochrome c + 2 H+
show the reaction diagram
-
-
-
-
(S)-lactate + 2 ferricytochrome c
pyruvate + 2 ferrocytochrome c + 2 H+
show the reaction diagram
-
-
-
-
(S)-lactate + 2 ferricytochrome c
pyruvate + 2 ferrocytochrome c + 2 H+
show the reaction diagram
-
-
-
-
(S)-lactate + 2 ferricytochrome c
pyruvate + 2 ferrocytochrome c + 2 H+
show the reaction diagram
-
-
-
-
(S)-lactate + 2 ferricytochrome c
pyruvate + 2 ferrocytochrome c + 2 H+
show the reaction diagram
-
-
-
-
(S)-lactate + 2 ferricytochrome c
pyruvate + 2 ferrocytochrome c + 2 H+
show the reaction diagram
-
-
-
-
(S)-lactate + 2 ferricytochrome c
pyruvate + 2 ferrocytochrome c + 2 H+
show the reaction diagram
-
-
-
-
(S)-lactate + 2 ferricytochrome c
pyruvate + 2 ferrocytochrome c + 2 H+
show the reaction diagram
-
-
-
-
(S)-lactate + 2 ferricytochrome c
pyruvate + 2 ferrocytochrome c + 2 H+
show the reaction diagram
-
-
-
-
(S)-lactate + 2 ferricytochrome c
pyruvate + 2 ferrocytochrome c + 2 H+
show the reaction diagram
-
-
-
-
(S)-lactate + 2 ferricytochrome c
pyruvate + 2 ferrocytochrome c + 2 H+
show the reaction diagram
-
-
-
-
(S)-lactate + 2 ferricytochrome c
pyruvate + 2 ferrocytochrome c + 2 H+
show the reaction diagram
-
-
-
-
(S)-lactate + 2 ferricytochrome c
pyruvate + 2 ferrocytochrome c + 2 H+
show the reaction diagram
-
-
-
-
(S)-lactate + 2 ferricytochrome c
pyruvate + 2 ferrocytochrome c + 2 H+
show the reaction diagram
-
-
-
-
(S)-lactate + 2 ferricytochrome c
pyruvate + 2 ferrocytochrome c + 2 H+
show the reaction diagram
-
-
-
-
(S)-lactate + 2 ferricytochrome c
pyruvate + 2 ferrocytochrome c + 2 H+
show the reaction diagram
P00175
-
-
-
(S)-lactate + 2 ferricytochrome c
pyruvate + 2 ferrocytochrome c + 2 H+
show the reaction diagram
-
-
-
-
?
(S)-lactate + 2 ferricytochrome c
pyruvate + 2 ferrocytochrome c + 2 H+
show the reaction diagram
-
-
-
r
(S)-lactate + 2 ferricytochrome c
pyruvate + 2 ferrocytochrome c + 2 H+
show the reaction diagram
-
-
-
r
(S)-lactate + 2 ferricytochrome c
pyruvate + 2 ferrocytochrome c + 2 H+
show the reaction diagram
-
-
-
-
?
(S)-lactate + 2 ferricytochrome c
pyruvate + 2 ferrocytochrome c + 2 H+
show the reaction diagram
-
-
-
-
(S)-lactate + 2 ferricytochrome c
pyruvate + 2 ferrocytochrome c + 2 H+
show the reaction diagram
-
-
-
-
(S)-lactate + 2 ferricytochrome c
pyruvate + 2 ferrocytochrome c + 2 H+
show the reaction diagram
-
-
-
-
(S)-lactate + 2 ferricytochrome c
pyruvate + 2 ferrocytochrome c + 2 H+
show the reaction diagram
-
-
-
-
(S)-lactate + 2 ferricytochrome c
pyruvate + 2 ferrocytochrome c + 2 H+
show the reaction diagram
-
-
-
-
(S)-lactate + 2 ferricytochrome c
pyruvate + 2 ferrocytochrome c + 2 H+
show the reaction diagram
-
-
-
r
(S)-lactate + 2 ferricytochrome c
pyruvate + 2 ferrocytochrome c + 2 H+
show the reaction diagram
-
-
-
-
?
(S)-lactate + 2 ferricytochrome c
pyruvate + 2 ferrocytochrome c + 2 H+
show the reaction diagram
-
D-isomer not oxidized
-
-
(S)-lactate + 2 ferricytochrome c
pyruvate + 2 ferrocytochrome c + 2 H+
show the reaction diagram
-
D-isomer not oxidized
-
?
(S)-lactate + 2 ferricytochrome c
pyruvate + 2 ferrocytochrome c + 2 H+
show the reaction diagram
-
can feed electrons to respiratory chain at the level of cytochrome c
-
-
(S)-lactate + 2 ferricytochrome c
pyruvate + 2 ferrocytochrome c + 2 H+
show the reaction diagram
-
can feed electrons to respiratory chain at the level of cytochrome c
-
-
(S)-lactate + 2 ferricytochrome c
pyruvate + 2 ferrocytochrome c + 2 H+
show the reaction diagram
-
can feed electrons to respiratory chain at the level of cytochrome c
-
-
(S)-lactate + 2 ferricytochrome c
pyruvate + 2 ferrocytochrome c + 2 H+
show the reaction diagram
-
can feed electrons to respiratory chain at the level of cytochrome c
-
-
(S)-lactate + 2 ferricytochrome c
pyruvate + 2 ferrocytochrome c + 2 H+
show the reaction diagram
-
can feed electrons to respiratory chain at the level of cytochrome c
-
-
(S)-lactate + 2 ferricytochrome c
pyruvate + 2 ferrocytochrome c + 2 H+
show the reaction diagram
-
can feed electrons to respiratory chain at the level of cytochrome c
-
-
(S)-lactate + 2 ferricytochrome c
pyruvate + 2 ferrocytochrome c + 2 H+
show the reaction diagram
-
can feed electrons to respiratory chain at the level of cytochrome c
-
-
(S)-lactate + 2 ferricytochrome c
pyruvate + 2 ferrocytochrome c + 2 H+
show the reaction diagram
-
can feed electrons to respiratory chain at the level of cytochrome c
-
-
(S)-lactate + 2 ferricytochrome c
pyruvate + 2 ferrocytochrome c + 2 H+
show the reaction diagram
-
can feed electrons to respiratory chain at the level of cytochrome c
-
-
(S)-lactate + 2 ferricytochrome c
pyruvate + 2 ferrocytochrome c + 2 H+
show the reaction diagram
-
can feed electrons to respiratory chain at the level of cytochrome c
-
-
(S)-lactate + 2 ferricytochrome c
pyruvate + 2 ferrocytochrome c + 2 H+
show the reaction diagram
-
can feed electrons to respiratory chain at the level of cytochrome c
-
-
(S)-lactate + 2 ferricytochrome c
pyruvate + 2 ferrocytochrome c + 2 H+
show the reaction diagram
-
can feed electrons to respiratory chain at the level of cytochrome c
-
-
(S)-lactate + 2 ferricytochrome c
pyruvate + 2 ferrocytochrome c + 2 H+
show the reaction diagram
-
can feed electrons to respiratory chain at the level of cytochrome c
-
-
(S)-lactate + 2 ferricytochrome c
pyruvate + 2 ferrocytochrome c + 2 H+
show the reaction diagram
-
can feed electrons to respiratory chain at the level of cytochrome c
-
-
(S)-lactate + 2 ferricytochrome c
pyruvate + 2 ferrocytochrome c + 2 H+
show the reaction diagram
-
can feed electrons to respiratory chain at the level of cytochrome c
-
-
(S)-lactate + 2 ferricytochrome c
pyruvate + 2 ferrocytochrome c + 2 H+
show the reaction diagram
-
can feed electrons to respiratory chain at the level of cytochrome c
-
-
(S)-lactate + 2 ferricytochrome c
pyruvate + 2 ferrocytochrome c + 2 H+
show the reaction diagram
-
can feed electrons to respiratory chain at the level of cytochrome c
-
-
(S)-lactate + 2 ferricytochrome c
pyruvate + 2 ferrocytochrome c + 2 H+
show the reaction diagram
P00175
can feed electrons to respiratory chain at the level of cytochrome c
-
-
(S)-lactate + 2 ferricytochrome c
pyruvate + 2 ferrocytochrome c + 2 H+
show the reaction diagram
-
can feed electrons to respiratory chain at the level of cytochrome c
-
r
(S)-lactate + 2 ferricytochrome c
pyruvate + 2 ferrocytochrome c + 2 H+
show the reaction diagram
-
can feed electrons to respiratory chain at the level of cytochrome c
-
r
(S)-lactate + 2 ferricytochrome c
pyruvate + 2 ferrocytochrome c + 2 H+
show the reaction diagram
-
replaces activity of EC 1.1.1.27 in limited substrate conditions
-
?
(S)-lactate + 2 ferricytochrome c
pyruvate + 2 ferrocytochrome c + 2 H+
show the reaction diagram
-
kcat/KM is 24000 fold lower with the recombinantly expressed flavocytochrome b2 flavin-binding domain compared to wild-type enzyme
-
-
?
(S)-lactate + 2 ferricytochrome c
pyruvate + 2 ferrocytochrome c + 2 H+
show the reaction diagram
Ogataea angusta C-105
-
-
-
-
?
(S)-lactate + 2 potassium ferricyanide
pyruvate + 2 reduced potassium ferricyanide + 2 H+
show the reaction diagram
-
-
-
-
?
(S)-lactate + cytochrome c
pyruvate + oxidized cytochrome c
show the reaction diagram
-
-
-
-
-
(S)-lactate + cytochrome c
pyruvate + oxidized cytochrome c
show the reaction diagram
-
-
-
-
?
(S)-lactate + cytochrome c
pyruvate + oxidized cytochrome c
show the reaction diagram
-
-
-
-
-
(S)-lactate + K3[Fe(CN)6]
?
show the reaction diagram
Ogataea angusta, Ogataea angusta C-105
-
-
-
-
?
(S)-lactate + potassium ferricyanide
pyruvate + potassium ferrocyanide
show the reaction diagram
-
-
-
-
?
(S)-mandelate + ferricytochrome c
hydroxy(phenyl)acetate + ferrocytochrome c
show the reaction diagram
-
-
-
-
-
(S)-mandelate + ferricytochrome c
hydroxy(phenyl)acetate + ferrocytochrome c
show the reaction diagram
-
traces of activity with wild type enzyme, significant activity with A198G/L230G double mutant
-
-
?
(S)-phenyllactate + ferricytochrome c
phenylpyruvate + ferrocytochrome c
show the reaction diagram
-
-
-
-
?
glycolate + ferricytochrome c
glyoxylate + ferrocytochrome c
show the reaction diagram
-
very poor substrate
-
-
?
L-lactate + ferricyanide
pyruvate + ferrocyanide + H+
show the reaction diagram
-
-
-
-
-
L-lactate + ferricyanide
pyruvate + ferrocyanide + H+
show the reaction diagram
-
-
-
-
?
L-lactate + ferricyanide
pyruvate + ferrocyanide + H+
show the reaction diagram
-
-
-
-
-
L-lactate + ferricytochrome c
pyruvate + ferrocytochrome c + H+
show the reaction diagram
-
-
-
-
?
L-lactate + ferricytochrome c
pyruvate + ferrocytochrome c + H+
show the reaction diagram
-
-
-
-
?
L-lactate + ferricytochrome c
pyruvate + ferrocytochrome c + H+
show the reaction diagram
-
-
-
-
?
L-lactate + ferricytochrome c
pyruvate + ferrocytochrome c + H+
show the reaction diagram
-
flavocytochrome b2, i.e. L-lactate cytochrome c oxidoreductase, catalyzes L-lactate oxidation at the expense of cytochrome c in the mitochondrial intermembrane space in yeast and enables the latter to grow on lactate as the sole carbon source
-
-
?
L-lactate + ferricytochrome c
pyruvate + ferrocytochrome c + H+
show the reaction diagram
-
molecular dynamics studies on active-site models of flavocytochrome b2 in complex with the substrate for analysis of the mechanism of the enzyme-catalyzed L-lactate oxidation reaction, overview. In the calculated enzyme-substrate model complex, the l-lactate alpha-OH hydrogen is hydrogen bonded to the activesite base H373 Ne, whereas the Halpha is directed towards flavin N5, suggesting that the reaction is initiated by a-OH proton abstraction
-
-
?
L-lactate + ferricytochrome c
pyruvate + ferrocytochrome c + H+
show the reaction diagram
Saccharomyces cerevisiae D273-10B/A1
-
-
-
-
?
L-lactate + O2
? + superoxide anion
show the reaction diagram
-
the FDH domain reacts slowly with oxygen with formation of superoxide anion when separated from its natural electron acceptor, whether isolated or included in the holoenzyme
-
-
?
L-lactate + potassium ferricyanide
pyruvate + potassium ferrocyanide + H+
show the reaction diagram
-
-
-
-
?
phenyllactate + ferricytochrome c
phenylpyruvate + ferrocytochrome c
show the reaction diagram
-
-
-
-
?
L-lactate + potassium ferricyanide
pyruvate + potassium ferrocyanide + H+
show the reaction diagram
-
-
-
-
?
additional information
?
-
-
2,6-dichlorophenolindophenol, ferricyanide and cytochrome c but not oxygen can serve as electron acceptor, rate of reduction with cytochrome c much slower than with the other possible acceptors, glycolate and L-malate do not serve as substrates
-
-
-
additional information
?
-
-
ferricyanide used as electron acceptor
-
-
-
additional information
?
-
P00175
ferricyanide used as electron acceptor
-
-
-
additional information
?
-
-
ferricyanide used as electron acceptor
-
-
-
additional information
?
-
-
2,6-dichloroindophenol, ferricyanide, methylene blue, 1,2-naphthoquinone and cytochrome c can serve as electron acceptors
-
-
-
additional information
?
-
-
2,6-dichlorophenolindophenol used as electron acceptor, 10times more sensitive than ferricyanide
-
-
-
additional information
?
-
-
electron acceptors other than ferricytochrome c used
-
-
-
additional information
?
-
P00175
electron acceptors other than ferricytochrome c used
-
-
-
additional information
?
-
-
electron acceptors other than ferricytochrome c used
-
-
-
additional information
?
-
-
electron acceptors other than ferricytochrome c used
-
-
-
additional information
?
-
-
during the catalytic cycle, electrons are transferred one by one from the reduced flavin to heme b2 in the same subunit
-
-
-
additional information
?
-
-
the role of the flavin mononucleotide-ribityl chain 2'OH group in maintaining the conserved K349 in a geometry favoring flavin reduction, of an active site water molecule belonging to a S371-Wat-D282-H373 hydrogen-bonded chain, which modulates the reactivity of the key catalytic histidine, and of the flavin C4a-C10a locus in facilitating proton transfer from the substrate to the active-site base, favoring the initial step of the lactate dehydrogenation reaction
-
-
-
additional information
?
-
-
chromate-reducing ability of enzyme-overproducing recombinant cells, the highest chromate-reducing activity of the cells is achieved in the presence of 2,6-dichlorophenolindophenol, the enzyme has absolute specificity for L-lactate, yet is non-selective with respect to its electron acceptor
-
-
-
additional information
?
-
Ogataea angusta C-105
-
chromate-reducing ability of enzyme-overproducing recombinant cells, the highest chromate-reducing activity of the cells is achieved in the presence of 2,6-dichlorophenolindophenol, the enzyme has absolute specificity for L-lactate, yet is non-selective with respect to its electron acceptor
-
-
-
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 + 2 ferricytochrome c
pyruvate + 2 ferrocytochrome c + 2 H+
show the reaction diagram
-
-
-
-
?
(S)-lactate + 2 ferricytochrome c
pyruvate + 2 ferrocytochrome c + 2 H+
show the reaction diagram
-
-
-
-
?
(S)-lactate + 2 ferricytochrome c
pyruvate + 2 ferrocytochrome c + 2 H+
show the reaction diagram
-
-
-
-
(S)-lactate + 2 ferricytochrome c
pyruvate + 2 ferrocytochrome c + 2 H+
show the reaction diagram
-
-
-
-
(S)-lactate + 2 ferricytochrome c
pyruvate + 2 ferrocytochrome c + 2 H+
show the reaction diagram
-
-
-
-
(S)-lactate + 2 ferricytochrome c
pyruvate + 2 ferrocytochrome c + 2 H+
show the reaction diagram
-
-
-
-
(S)-lactate + 2 ferricytochrome c
pyruvate + 2 ferrocytochrome c + 2 H+
show the reaction diagram
-
-
-
-
(S)-lactate + 2 ferricytochrome c
pyruvate + 2 ferrocytochrome c + 2 H+
show the reaction diagram
-
-
-
r
(S)-lactate + 2 ferricytochrome c
pyruvate + 2 ferrocytochrome c + 2 H+
show the reaction diagram
-
-
-
-
?
(S)-lactate + 2 ferricytochrome c
pyruvate + 2 ferrocytochrome c + 2 H+
show the reaction diagram
-
can feed electrons to respiratory chain at the level of cytochrome c
-
-
(S)-lactate + 2 ferricytochrome c
pyruvate + 2 ferrocytochrome c + 2 H+
show the reaction diagram
-
can feed electrons to respiratory chain at the level of cytochrome c
-
-
(S)-lactate + 2 ferricytochrome c
pyruvate + 2 ferrocytochrome c + 2 H+
show the reaction diagram
-
can feed electrons to respiratory chain at the level of cytochrome c
-
-
(S)-lactate + 2 ferricytochrome c
pyruvate + 2 ferrocytochrome c + 2 H+
show the reaction diagram
-
can feed electrons to respiratory chain at the level of cytochrome c
-
-
(S)-lactate + 2 ferricytochrome c
pyruvate + 2 ferrocytochrome c + 2 H+
show the reaction diagram
-
can feed electrons to respiratory chain at the level of cytochrome c
-
-
(S)-lactate + 2 ferricytochrome c
pyruvate + 2 ferrocytochrome c + 2 H+
show the reaction diagram
-
can feed electrons to respiratory chain at the level of cytochrome c
-
-
(S)-lactate + 2 ferricytochrome c
pyruvate + 2 ferrocytochrome c + 2 H+
show the reaction diagram
-
can feed electrons to respiratory chain at the level of cytochrome c
-
-
(S)-lactate + 2 ferricytochrome c
pyruvate + 2 ferrocytochrome c + 2 H+
show the reaction diagram
-
can feed electrons to respiratory chain at the level of cytochrome c
-
-
(S)-lactate + 2 ferricytochrome c
pyruvate + 2 ferrocytochrome c + 2 H+
show the reaction diagram
-
can feed electrons to respiratory chain at the level of cytochrome c
-
-
(S)-lactate + 2 ferricytochrome c
pyruvate + 2 ferrocytochrome c + 2 H+
show the reaction diagram
-
can feed electrons to respiratory chain at the level of cytochrome c
-
-
(S)-lactate + 2 ferricytochrome c
pyruvate + 2 ferrocytochrome c + 2 H+
show the reaction diagram
-
can feed electrons to respiratory chain at the level of cytochrome c
-
-
(S)-lactate + 2 ferricytochrome c
pyruvate + 2 ferrocytochrome c + 2 H+
show the reaction diagram
-
can feed electrons to respiratory chain at the level of cytochrome c
-
-
(S)-lactate + 2 ferricytochrome c
pyruvate + 2 ferrocytochrome c + 2 H+
show the reaction diagram
-
can feed electrons to respiratory chain at the level of cytochrome c
-
-
(S)-lactate + 2 ferricytochrome c
pyruvate + 2 ferrocytochrome c + 2 H+
show the reaction diagram
-
can feed electrons to respiratory chain at the level of cytochrome c
-
-
(S)-lactate + 2 ferricytochrome c
pyruvate + 2 ferrocytochrome c + 2 H+
show the reaction diagram
-
can feed electrons to respiratory chain at the level of cytochrome c
-
-
(S)-lactate + 2 ferricytochrome c
pyruvate + 2 ferrocytochrome c + 2 H+
show the reaction diagram
-
can feed electrons to respiratory chain at the level of cytochrome c
-
-
(S)-lactate + 2 ferricytochrome c
pyruvate + 2 ferrocytochrome c + 2 H+
show the reaction diagram
-
can feed electrons to respiratory chain at the level of cytochrome c
-
-
(S)-lactate + 2 ferricytochrome c
pyruvate + 2 ferrocytochrome c + 2 H+
show the reaction diagram
P00175
can feed electrons to respiratory chain at the level of cytochrome c
-
-
(S)-lactate + 2 ferricytochrome c
pyruvate + 2 ferrocytochrome c + 2 H+
show the reaction diagram
-
can feed electrons to respiratory chain at the level of cytochrome c
-
r
(S)-lactate + 2 ferricytochrome c
pyruvate + 2 ferrocytochrome c + 2 H+
show the reaction diagram
-
can feed electrons to respiratory chain at the level of cytochrome c
-
r
(S)-lactate + 2 ferricytochrome c
pyruvate + 2 ferrocytochrome c + 2 H+
show the reaction diagram
-
replaces activity of EC 1.1.1.27 in limited substrate conditions
-
?
(S)-lactate + 2 ferricytochrome c
pyruvate + 2 ferrocytochrome c + 2 H+
show the reaction diagram
Ogataea angusta C-105
-
-
-
-
?
L-lactate + ferricytochrome c
pyruvate + ferrocytochrome c + H+
show the reaction diagram
-
-
-
-
?
L-lactate + ferricytochrome c
pyruvate + ferrocytochrome c + H+
show the reaction diagram
-
-
-
-
?
L-lactate + ferricytochrome c
pyruvate + ferrocytochrome c + H+
show the reaction diagram
-
-
-
-
?
L-lactate + ferricytochrome c
pyruvate + ferrocytochrome c + H+
show the reaction diagram
-
flavocytochrome b2, i.e. L-lactate cytochrome c oxidoreductase, catalyzes L-lactate oxidation at the expense of cytochrome c in the mitochondrial intermembrane space in yeast and enables the latter to grow on lactate as the sole carbon source
-
-
?
L-lactate + ferricytochrome c
pyruvate + ferrocytochrome c + H+
show the reaction diagram
Saccharomyces cerevisiae D273-10B/A1
-
-
-
-
?
additional information
?
-
Ogataea angusta, Ogataea angusta C-105
-
chromate-reducing ability of enzyme-overproducing recombinant cells, the highest chromate-reducing activity of the cells is achieved in the presence of 2,6-dichlorophenolindophenol
-
-
-
COFACTOR
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
2,6-dichlorophenolindophenol
-
-
cytochrome
-
each subunit of the tetramer is composed of two domains, one binding a heme and the other an FMN prosthetic group. The cytochrome domain consists of residues 1 to 99
-
cytochrome c
-
-
cytochrome c
-
the flavoenzyme possesses flavocytochrome b2 subunits, that each consists of an N-terminal cytochrome domain and a C-terminal flavodehydrogenase, FDH, domain
cytochrome c
-
-
FAD
-
the flavoenzyme possesses flavocytochrome b2 subunits, that each consists of an N-terminal cytochrome domain and a C-terminal flavodehydrogenase, FDH, domain
ferricytochrome c
-
-
ferricytochrome c
-
-
flavocytochrome c
-
-
-
FMN
-
one FMN per subunit
FMN
-
the C-terminal domain of the enzyme contains FMN
FMN
-
each subunit of the tetramer is composed of two domains, one binding a heme and the other an FMN prosthetic group. The flavin binding domain contains a parallel beta8alpha8 barrel structure and is composed of residues 100 to 486. The FMN moiety, which is located at the C-terminal end of the central beta-barrel, is mostly sequestered from solvent. It forms hydrogen bond interactions with main- and side-chain atoms from six of the eight beta-strands. The interaction of Lys349 with atoms N-1 and O-2 of the flavin ring is probably responsible for stabilization of the anionic form of the flavin semiquinone and hydroquinone and enhancing the reactivity of atom N-5 toward sulfite. The binding of pyruvate at the active site in subunit 2 is stabilized by interaction of its carboxylate group with the side-chain atoms of Arg376 and Tyr143. Residues His373 and Tyr254 interact with the keto-oxygen atom and are involved in catalysis. In contrast, four water molecules occupy the substrate-binding site in subunit 1 and Tyr143 forms a hydrogen bond to the ordered heme propionate group. Otherwise the two flavin-binding domains are identical within experimental error
FMN
-
flavohemoprotein, in the crystal structure, FMN and heme are face to face, and appear to be in a suitable orientation and at a suitable distance for exchanging electrons
heme
-
one heme per subunit
heme
-
the N-terminal domain of the enzyme contains protohaem IX
heme
-
the flavoenzyme possesses flavocytochrome b2 subunits, that each consists of an N-terminal cytochrome domain and a C-terminal flavodehydrogenase, FDH, domain
heme
-
each enzyme subunit contains a cytochrome b5-like heme domain
heme
-
flavohemoprotein, in the crystal structure, FMN and heme are face to face, and appear to be in a suitable orientation and at a suitable distance for exchanging electrons. But in one subunit out of two, the heme domain is disordered and invisible. The heme domains are mobile in solution
additional information
-
electron acceptors other than ferricytochrome c used; NAD+ and NADP+ cannot substitute, 2,6-dichlorophenolindophenol, ferricyanide and cytochrome c but not oxygen can serve as electron acceptor, rate of reduction with cytochrome c much slower than with the other possible acceptors
-
additional information
-
2,6-dichloroindophenol, ferricyanide, methylene blue, 1,2-naphthoquinone and cytochrome c can serve as electron acceptors; electron acceptors other than ferricytochrome c used
-
additional information
-
electron acceptors other than ferricytochrome c used
-
additional information
-
electron acceptors other than ferricytochrome c used; ferricyanide used as electron acceptor
-
additional information
-
2,6-dichlorophenolindophenol used as electron acceptor, 10-fold more sensitive than ferricyanide; electron acceptors other than ferricytochrome c used
-
additional information
-
electron acceptors other than ferricytochrome c used
-
additional information
-
electron acceptors other than ferricytochrome c used; ferricyanide used as electron acceptor
-
additional information
-
the enzyme is active with potassium ferricyanide
-
additional information
-
binding studies and models of the complex between cytochrome c and Fcb2, E63 is involved in the interaction with cyt. c, overview. Cytochrome c mutants E63K/D72K and E63K/D72K show reduced activity, but neither the K296M nor the Y97F mutation show significantly alteration of the enzyme kinetics. Effect of mutations on heme and cytochrome binding, overview
-
METALS and IONS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
Fe2+
-
bound to enzyme
Fe2+
-
heme cofactor
INHIBITORS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
ethane nitronate
-
competitive
ferricyanide
-
-
L-lactate
-
inhibition by excess of substrate, no inhibition with Saccharomyces cerevisiae enzyme
L-lactate
-
60% inhibition at 10 mM, no inhibition with cleaved form of enzyme
L-lactate
-
40% inhibition at 60 mM
L-Mandelate
-
competitive
malate
-
inhibitory to isoform with activity maximum at pH 9.5
oxalate
-
complete inhibition, competitive
p-mercuribenzoate
-
-
Propionate
-
competitive
pyruvate
-
noncompetitive at low concentrations, uncompetitve at high concentrations
pyruvate
-
non-competitive
pyruvate
-
competitive at low concentrations, non-competitve at high concentrations
zinc-substituted cytochrome c
-
-
-
malate
-
competitive inhibition
additional information
-
no inhibition with nitroethane
-
additional information
-
enzyme is inhibited by high substrate concentrations
-
KM VALUE [mM]
KM VALUE [mM] Maximum
SUBSTRATE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
0.16
-
(S)-Lactate
-
pH 7.0, 25C, wild type
0.19
-
(S)-Lactate
-
pH 7.5, 25C, Tris-HCl buffer, mutant Y143F, with heme
0.28
-
(S)-Lactate
-
pH 7.0, 25C, mutant Y254F
0.4
-
(S)-Lactate
-
pH 7.0, 5C, phosphate buffer, mutant Y143F, with heme
0.53
-
(S)-Lactate
-
pH 7.5, 25C, Tris-HCl buffer, wild-type enzyme, with heme
0.54
-
(S)-Lactate
-
pH 7.0, 5C, phosphate buffer, wild-type enzyme, with heme
0.84
-
(S)-Lactate
-
pH 7.5, 25C, Tris-HCl buffer, wild-type enzyme, with FMN
0.89
-
(S)-Lactate
-
pH 7.0, 5C, phosphate buffer, wild-type enzyme, with FMN
2.09
-
(S)-Lactate
-
pH 7.0, 20C
2.5
5
(S)-Lactate
-
pH 7.0, 5C, phosphate buffer, mutant Y143F, with FMN
2.6
-
(S)-Lactate
-
pH 8.0, temperature not specified in the publication
2.81
-
(S)-Lactate
-
pH 7.5, 25C, Tris-HCl buffer, mutant Y143F, with FMN
0.02
-
cytochrome c
-
pH 7.0, 30C, flavin turnover in pre-steady-state flavin reduction, mutant P44A
0.032
-
cytochrome c
-
pH 7.0, 30C, flavin turnover in pre-steady-state flavin reduction, mutant A67Q
0.051
-
cytochrome c
-
pH 7.0, 30C, flavin turnover in pre-steady-state flavin reduction, mutant L65A
0.061
-
cytochrome c
-
pH 7.0, 30C, flavin turnover in pre-steady-state flavin reduction, mutant P64R
0.065
-
cytochrome c
-
pH 7.0, 30C, flavin turnover in pre-steady-state flavin reduction, mutant F39A
0.069
-
cytochrome c
-
pH 7.0, 30C, flavin turnover in pre-steady-state flavin reduction, mutant P64Q
0.085
-
cytochrome c
-
pH 7.0, 30C, flavin turnover in pre-steady-state flavin reduction, mutant A67L
0.092
-
cytochrome c
-
pH 7.0, 30C, flavin turnover in pre-steady-state flavin reduction, mutant N69K
0.097
-
cytochrome c
-
pH 7.0, 30C, flavin turnover in pre-steady-state flavin reduction, mutant D72A
0.105
-
cytochrome c
-
pH 7.0, 30C, flavin turnover in pre-steady-state flavin reduction, mutant V70M
0.109
-
cytochrome c
-
pH 7.0, 30C, flavin turnover in pre-steady-state flavin reduction, mutant K73A
0.113
-
cytochrome c
-
pH 7.0, 30C, flavin turnover in pre-steady-state flavin reduction, mutant F39A
0.119
-
cytochrome c
-
pH 7.0, 30C, flavin turnover in pre-steady-state flavin reduction, mutant E63K
0.131
-
cytochrome c
-
pH 7.0, 30C, flavin turnover in pre-steady-state flavin reduction, wild-type enzyme
0.7
-
cytochrome c
-
Y254L mutant
2.4
-
cytochrome c
-
Y254F mutant
131
-
cytochrome c
-
wild type enzyme
0.03
-
ferricyanide
-
Y254L mutant
0.04
-
ferricyanide
-
Y254F mutant
0.07
-
ferricyanide
-
wild type enzyme
0.07
-
ferricyanide
-
pH 5.5, isoform with activity maximum at pH 5.5
0.138
-
ferricyanide
-
pH 7.5, isoform with activity maximum at pH 7.5
0.27
-
ferricyanide
-
pH 9.5, isoform with activity maximum at pH 9.5
0.0015
-
ferricytochrome c
-
pH 7.5, 25C, Tris-HCl buffer, mutant Y143F
0.01
-
ferricytochrome c
-
pH 7.5, 25C, wild-type enzyme
0.01
-
ferricytochrome c
-
pH 7.5, 25C, Tris-HCl buffer, wild-type enzyme
0.023
-
ferricytochrome c
-
pH 7.5, 25C, recombinantly expressed flavocytochrome b2 flavin-binding domain
0.045
-
ferricytochrome c
-
pH 7.0, 5C, phosphate buffer, wild-type enzyme
0.121
-
ferricytochrome c
-
pH 7.0, 30C, phosphate buffer, mutant Y143F
0.131
-
ferricytochrome c
-
pH 7.0, 30C, phosphate buffer, wild-type enzyme; pH 7.0, 5C, phosphate buffer, mutant Y143F
0.0001
-
L-lactate
-
Y254L mutant enzyme with 2,6-dichloroindophenol as electron acceptor
0.037
-
L-lactate
-
wild type enzyme with 2,6-dichloroindophenol as electron acceptor
0.04
-
L-lactate
-
Y143F mutant enzyme with 2,6-dichloroindophenol as electron acceptor
0.13
-
L-lactate
-
H373Q mutant enzyme with 2,6-dichloroindophenol as electron acceptor
0.16
-
L-lactate
-
wild-type, intact protein
0.23
-
L-lactate
-
Y143F mutant with cytochrome c as electron acceptor
0.24
-
L-lactate
-
native enzyme with cytochrome c as electron acceptor
0.25
-
L-lactate
-
mutant H373Q, using flavin domain only
0.29
-
L-lactate
-
wild type enzyme
0.33
-
L-lactate
-
sensor based on the inital Hansenula polymorpha C-105 cells
0.34
-
L-lactate
-
Y254F mutant
0.35
-
L-lactate
-
Y254F mutant
0.36
-
L-lactate
-
wild-type, using flavin domain only
0.4
-
L-lactate
-
intact enzyme
0.4
-
L-lactate
-
A198G mutant enzyme
0.49
-
L-lactate
-
wild type enzyme
0.49
-
L-lactate
-
native enzyme mutant with ferricyanide as electron acceptor
0.49
-
L-lactate
-
wild type enzyme
0.5
-
L-lactate
-
wild type enzyme
0.53
-
L-lactate
-
Y254L mutant
0.6
-
L-lactate
-
L230A mutant enzyme
0.65
-
L-lactate
-
mutant H373Q, intact protein
0.66
-
L-lactate
-
FDH domain, reaction conditions: 13 mM ferricyanide, 100 mM phosphate buffer, 1 mM EDTA, pH 7, 30C; FDH domain, reaction conditions: 13 mM ferricyanide, 100 mM phosphate buffer, 1 mM EDTA, pH 7.5, 25C
0.86
-
L-lactate
-
FDH domain, reaction conditions: 13 mM ferricyanide, 10 mM Tris/HCl buffer, 0.1 M NaCl, pH 7.5, 25C
0.89
-
L-lactate
-
stopped-flow kinetic parameters for flavin reduction by L-lactate using holo-enzyme, reaction conditions: 100 mM phosphate buffer, 1 mM EDTA, pH 7, 5C, in the absence of ferrocyanide
0.9
-
L-lactate
-
influence of anions (200 mM phsophate) on the wild-type steady-state kinetic. Reaction conditions: 100 mM phosphate buffer, 1 mM EDTA, pH 7, 5C, 1.5 mM ferricyanide and variable L-lactate concentration
0.94
-
L-lactate
-
influence of anions (400 mM KBr) on the wild-type steady-state kinetic. Reaction conditions: 100 mM phosphate buffer, 1 mM EDTA, pH 7, 5C, 1.5 mM ferricyanide and variable L-lactate concentration
1.03
-
L-lactate
-
stopped-flow kinetic parameters for flavin reduction by L-lactate using FDH domain, reaction conditions: 100 mM phosphate buffer, 1 mM EDTA, pH 7, 5C, in the absence of ferrocyanide
1.18
-
L-lactate
-
FDH domain, reaction conditions: 13 mM ferricyanide, 100 mM phosphate buffer, 1 mM EDTA, pH 7, 5C
1.5
-
L-lactate
-
influence of anions (400 mM KCl) on the wild-type steady-state kinetic. Reaction conditions: 100 mM phosphate buffer, 1 mM EDTA, pH 7, 5C, 1.5 mM ferricyanide and variable L-lactate concentration
1.6
-
L-lactate
-
cleaved enzyme
2.3
-
L-lactate
-
influence of anions (400 mM KCl) on the FDH domain steady-state kinetic. Reaction conditions: 100 mM phosphate buffer, 1 mM EDTA, pH 7, 5C, constant 10 mM ferricyanide and variable L-lactate concentration
2.7
-
L-lactate
-
influence of anions (400 mM potassium acetate) on the wild-type steady-state kinetic. Reaction conditions: 100 mM phosphate buffer, 1 mM EDTA, pH 7, 5C, 1.5 mM ferricyanide and variable L-lactate concentration
2.8
-
L-lactate
-
influence of anions (300 mM phsophate) on the FDH domain steady-state kinetic. Reaction conditions: 100 mM phosphate buffer, 1 mM EDTA, pH 7, 5C, constant 10 mM ferricyanide and variable L-lactate concentration
2.9
-
L-lactate
-
Y143F mutant with ferricyanide as electron acceptor
3
-
L-lactate
-
with ferricyanide as electron acceptor
3.02
-
L-lactate
-
biosensor based on recombinant yeast FCb2cells
3.85
-
L-lactate
-
2,6-dichlorophenolindopenol as electron acceptor
3.9
-
L-lactate
-
Y254F mutant
4
-
L-lactate
-
influence of anions (400 mM KBr) on the FDH domain steady-state kinetic. Reaction conditions: 100 mM phosphate buffer, 1 mM EDTA, pH 7, 5C, constant 10 mM ferricyanide and variable L-lactate concentration
4.6
-
L-lactate
-
influence of anions (400 mM potassium acetate) on the FDH domain steady-state kinetic. Reaction conditions: 100 mM phosphate buffer, 1 mM EDTA, pH 7, 5C, constant 10 mM ferricyanide and variable L-lactate concentration
5.8
-
L-lactate
-
influence of anions (400 mM KBr) on mutant R289K steady-state kinetic. Reaction conditions: 100 mM phosphate buffer, 1 mM EDTA, pH 7, 5C, 2 mM ferricyanide and variable L-lactate concentration
6.5
-
L-lactate
-
influence of anions (400 mM KCl) on mutant R289K steady-state kinetic. Reaction conditions: 100 mM phosphate buffer, 1 mM EDTA, pH 7, 5C, 2 mM ferricyanide and variable L-lactate concentration
7
-
L-lactate
-
mutant R289K steady-state kinetic. Reaction conditions: 100 mM phosphate buffer, 1 mM EDTA, pH 7, 5C, 2 mM ferricyanide and variable L-lactate concentration
8.7
-
L-lactate
-
influence of anions (200 mM phsophate) on mutant R289K steady-state kinetic. Reaction conditions: 100 mM phosphate buffer, 1 mM EDTA, pH 7, 5C, 2 mM ferricyanide and variable L-lactate concentration
8.9
-
L-lactate
-
wild type enzyme
9.2
-
L-lactate
-
influence of anions (400 mM potassium acetate) on mutant R289K steady-state kinetic. Reaction conditions: 100 mM phosphate buffer, 1 mM EDTA, pH 7, 5C, 2 mM ferricyanide and variable L-lactate concentration
38
-
L-lactate
-
A198G/L230A mutant enzyme
38
-
L-lactate
-
A198G/L230G mutant enzyme
0.22
-
L-Phenyllactate
-
wild type enzyme
0.084
-
Phenyllactate
-
Y254F mutant
0.4
-
Phenyllactate
-
wild type enzyme
6
-
potassium ferricyanide
-
pH 9.5, temperature not specified in the publication
1.43
-
L-Phenyllactate
-
R289K mutant enzyme
additional information
-
additional information
-
steady-state kinetic parameters and 2H kinetic isotope effects for the isolated flavin domain and intact, wild-type flavocytoochrome b2
-
additional information
-
additional information
-
stopped-flow, pre-steady-state and steady-state kinetics measuring electron transfer in artificial systems, redox potentials, overview
-
TURNOVER NUMBER [1/s]
TURNOVER NUMBER MAXIMUM[1/s]
SUBSTRATE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
13.2
-
(S)-Lactate
-
pH 7.0, 25C, mutant Y254F
372
-
(S)-Lactate
-
pH 7.0, 25C, wild type
6
-
cytochrome c
-
pH 7.0, 30C, flavin turnover in pre-steady-state flavin reduction, mutant A67Q
39
-
cytochrome c
-
pH 7.0, 30C, flavin turnover in pre-steady-state flavin reduction, mutant F39A
62
-
cytochrome c
-
pH 7.0, 30C, flavin turnover in pre-steady-state flavin reduction, mutant F39A
63
-
cytochrome c
-
pH 7.0, 30C, flavin turnover in pre-steady-state flavin reduction, mutant L65A
87
-
cytochrome c
-
pH 7.0, 30C, flavin turnover in pre-steady-state flavin reduction, mutant P44A
108
-
cytochrome c
-
pH 7.0, 30C, flavin turnover in pre-steady-state flavin reduction, mutant A67L
139
-
cytochrome c
-
pH 7.0, 30C, flavin turnover in pre-steady-state flavin reduction, mutant E63K
143
-
cytochrome c
-
pH 7.0, 30C, flavin turnover in pre-steady-state flavin reduction, mutant P64R
155
-
cytochrome c
-
pH 7.0, 30C, flavin turnover in pre-steady-state flavin reduction, wild-type enzyme
164
-
cytochrome c
-
pH 7.0, 30C, flavin turnover in pre-steady-state flavin reduction, mutant D72A
165
-
cytochrome c
-
pH 7.0, 30C, flavin turnover in pre-steady-state flavin reduction, mutant K73A
168
-
cytochrome c
-
pH 7.0, 30C, flavin turnover in pre-steady-state flavin reduction, mutant P64Q
171
-
cytochrome c
-
pH 7.0, 30C, flavin turnover in pre-steady-state flavin reduction, mutant V70M
184
-
cytochrome c
-
pH 7.0, 30C, flavin turnover in pre-steady-state flavin reduction, mutant N69K
185
-
FAD
-
pH 7.0, 30C, flavin turnover in pre-steady-state flavin reduction, mutant A67Q
190
-
FAD
-
pH 7.0, 30C, flavin turnover in pre-steady-state flavin reduction, mutant L65A
196
-
FAD
-
pH 7.0, 30C, flavin turnover in pre-steady-state flavin reduction, mutant F39A
207
-
FAD
-
pH 7.0, 30C, flavin turnover in pre-steady-state flavin reduction, mutant P64Q
208
-
FAD
-
pH 7.0, 30C, flavin turnover in pre-steady-state flavin reduction, mutant A67L
209
-
FAD
-
pH 7.0, 30C, flavin turnover in pre-steady-state flavin reduction, mutant F39A
211
-
FAD
-
pH 7.0, 30C, flavin turnover in pre-steady-state flavin reduction, mutant P44A
212
-
FAD
-
pH 7.0, 30C, flavin turnover in pre-steady-state flavin reduction, mutant P64R
214
-
FAD
-
pH 7.0, 30C, flavin turnover in pre-steady-state flavin reduction, mutant D72A; pH 7.0, 30C, flavin turnover in pre-steady-state flavin reduction, wild-type enzyme
218
-
FAD
-
pH 7.0, 30C, flavin turnover in pre-steady-state flavin reduction, mutant E63K; pH 7.0, 30C, flavin turnover in pre-steady-state flavin reduction, mutant N69K
223
-
FAD
-
pH 7.0, 30C, flavin turnover in pre-steady-state flavin reduction, mutant K73A
225
-
FAD
-
pH 7.0, 30C, flavin turnover in pre-steady-state flavin reduction, mutant V70M
0.011
-
ferricytochrome c
-
pH 7.5, 25C, Tris-HCl buffer, mutant Y143F
0.02
-
ferricytochrome c
-
pH 7.5, 25C, recombinantly expressed flavocytochrome b2 flavin-binding domain
0.103
-
ferricytochrome c
-
pH 7.5, 25C, Tris-HCl buffer, wild-type enzyme
20
-
ferricytochrome c
-
pH 7.0, 5C, phosphate buffer, mutant Y143F
61
-
ferricytochrome c
-
pH 7.0, 30C, phosphate buffer, mutant Y143F; pH 7.0, 5C, phosphate buffer, wild-type enzyme
155
-
ferricytochrome c
-
pH 7.0, 30C, phosphate buffer, wild-type enzyme
207
-
ferricytochrome c
-
pH 7.5, 25C, wild-type enzyme
0.2
-
Fluoropyruvate
-
Y254F mutant enzyme
2.8
-
Fluoropyruvate
-
wild type enzyme
0.031
-
L-lactate
-
mutant H373Q, intact protein
0.057
-
L-lactate
-
mutant H373Q, using flavin domain only
0.39
-
L-lactate
-
Y254L mutant enzyme
0.42
-
L-lactate
-
Y254L mutant
6.1
-
L-lactate
-
Y254F mutant enzyme
7.5
-
L-lactate
-
Y254F mutant
7.8
-
L-lactate
-
influence of anions (400 mM KBr) on mutant R289K steady-state kinetic. Reaction conditions: 100 mM phosphate buffer, 1 mM EDTA, pH 7, 5C, 2 mM ferricyanide and variable L-lactate concentration
8.6
-
L-lactate
-
mutant R289K steady-state kinetic. Reaction conditions: 100 mM phosphate buffer, 1 mM EDTA, pH 7, 5C, 2 mM ferricyanide and variable L-lactate concentration
8.8
-
L-lactate
-
influence of anions (400 mM potassium acetate) on mutant R289K steady-state kinetic. Reaction conditions: 100 mM phosphate buffer, 1 mM EDTA, pH 7, 5C, 2 mM ferricyanide and variable L-lactate concentration
9.2
-
L-lactate
-
influence of anions (200 mM phsophate) on mutant R289K steady-state kinetic. Reaction conditions: 100 mM phosphate buffer, 1 mM EDTA, pH 7, 5C, 2 mM ferricyanide and variable L-lactate concentration; influence of anions (400 mM KCl) on mutant R289K steady-state kinetic. Reaction conditions: 100 mM phosphate buffer, 1 mM EDTA, pH 7, 5C, 2 mM ferricyanide and variable L-lactate concentration
11
-
L-lactate
-
Y143F mutant with cytochrome c as electron acceptor
30
-
L-lactate
-
L230A mutant enzyme
41
-
L-lactate
-
A198G/L230A mutant enzyme
41
-
L-lactate
-
A198G/L230G mutant enzyme with ferricyanide as electron acceptor
45
-
L-lactate
-
influence of anions (400 mM KBr) on the FDH domain steady-state kinetic. Reaction conditions: 100 mM phosphate buffer, 1 mM EDTA, pH 7, 5C, constant 10 mM ferricyanide and variable L-lactate concentration
60
-
L-lactate
-
Y143F mutant enzyme
60
-
L-lactate
-
influence of anions (400 mM KCl) on the FDH domain steady-state kinetic. Reaction conditions: 100 mM phosphate buffer, 1 mM EDTA, pH 7, 5C, constant 10 mM ferricyanide and variable L-lactate concentration
61
-
L-lactate
-
influence of anions (400 mM KBr) on the wild-type steady-state kinetic. Reaction conditions: 100 mM phosphate buffer, 1 mM EDTA, pH 7, 5C, 1.5 mM ferricyanide and variable L-lactate concentration
71
-
L-lactate
-
influence of anions (200 mM phsophate) on the wild-type steady-state kinetic. Reaction conditions: 100 mM phosphate buffer, 1 mM EDTA, pH 7, 5C, 1.5 mM ferricyanide and variable L-lactate concentration
75
-
L-lactate
-
influence of anions (400 mM KCl) on the wild-type steady-state kinetic. Reaction conditions: 100 mM phosphate buffer, 1 mM EDTA, pH 7, 5C, 1.5 mM ferricyanide and variable L-lactate concentration
86
-
L-lactate
-
influence of anions (400 mM potassium acetate) on the FDH domain steady-state kinetic. Reaction conditions: 100 mM phosphate buffer, 1 mM EDTA, pH 7, 5C, constant 10 mM ferricyanide and variable L-lactate concentration
101
-
L-lactate
-
influence of anions (300 mM phsophate) on the FDH domain steady-state kinetic. Reaction conditions: 100 mM phosphate buffer, 1 mM EDTA, pH 7, 5C, constant 10 mM ferricyanide and variable L-lactate concentration
103
-
L-lactate
-
native enzyme with cytochrome c as electron acceptor
113
-
L-lactate
-
influence of anions (400 mM potassium acetate) on the wild-type steady-state kinetic. Reaction conditions: 100 mM phosphate buffer, 1 mM EDTA, pH 7, 5C, 1.5 mM ferricyanide and variable L-lactate concentration
117
-
L-lactate
-
holo-enzyme, reaction conditions: 1 or 2 mM ferricyanide, 100 mM phosphate buffer, 1mM EDTA, pH 7, 5C
133
-
L-lactate
-
FDH domain, reaction conditions: 13 mM ferricyanide, 100 mM phosphate buffer, 1 mM EDTA, pH 7, 5C
144
-
L-lactate
-
stopped-flow kinetic parameters for flavin reduction by L-lactate using holo-enzyme, reaction conditions: 100 mM phosphate buffer, 1 mM EDTA, pH 7, 5C, in the absence of ferrocyanide
149
-
L-lactate
-
stopped-flow kinetic parameters for flavin reduction by L-lactate using FDH domain, reaction conditions: 100 mM phosphate buffer, 1 mM EDTA, pH 7, 5C, in the absence of ferrocyanide
155
-
L-lactate
-
wild type enzyme
185
-
L-lactate
-
A198G mutant enzyme
200
-
L-lactate
-
wild-type, using flavin domain only
214
-
L-lactate
-
FDH domain, reaction conditions: 13 mM ferricyanide, 10 mM Tris/HCl buffer, 0.1 M NaCl, pH 7.5, 25C
240
-
L-lactate
-
FDH domain, reaction conditions: 13 mM ferricyanide, 100 mM phosphate buffer, 1 mM EDTA, pH 7.5, 25C
259
-
L-lactate
-
FDH domain, reaction conditions: 13 mM ferricyanide, 100 mM phosphate buffer, 1 mM EDTA, pH 7, 30C
270
-
L-lactate
-
wild type enzyme
372
-
L-lactate
-
wild-type, intact protein
400
-
L-lactate
-
wild type enzyme
400
-
L-lactate
-
native enzyme and Y143F mutant with ferricyanide as electron acceptor
400
-
L-lactate
-
wild type enzyme
473
-
L-lactate
-
wild type enzyme
0.02
-
L-Mandelate
-
wild type enzyme with ferricyanide as electron acceptor
0.29
-
Phenyllactate
-
Y254F mutant enzyme
16.5
-
Phenyllactate
-
wild type enzyme
8.5
-
L-Mandelate
-
A198G/L230G mutant enzyme with ferricyanide as electron acceptor
additional information
-
additional information
-
systematic determination of substrates with different chain length
-
additional information
-
additional information
-
systematic determination of turnover numbers for wild type enzyme and deletion mutants with different electron acceptors, significantly reduced activity for deletion mutants with cytochrome c as electron acceptor
-
additional information
-
additional information
-
systematic determination of activity of all mutants with L-mandelate and L-lactate as substrate
-
additional information
-
additional information
-
steady-state kinetic parameters and 2H kinetic isotope effects for the isolated flavin domain and intact, wild-type flavocytoochrome b2
-
additional information
-
additional information
-
second-order rate constant for cytochrome c reduction in the pre-steady-state determined by stopped-flow spectrophotometry
-
Ki VALUE [mM]
Ki VALUE [mM] Maximum
INHIBITOR
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
170
-
acetate
-
in pre-steady-state kinetic studies of the FDH domain reduction and reoxidation, acetate acts as a competitive inhibitor of L-lactate
450
-
chloride
-
in pre-steady-state kinetic studies of the FDH domain reduction and reoxidation, chloride acts as a competitive inhibitor of L-lactate
1.4
-
D-lactate
-
intact enzyme
6
-
D-lactate
-
cleaved enzyme
2.2
-
ethane nitronate
-
-
0.006
-
Ferrocytochrome
-
pH 7.5, 25C, mutant enzyme E91K
0.0088
-
Ferrocytochrome
-
pH 7.5, 25C, wild-type enzyme
0.0026
-
L-Mandelate
-
-
13
-
malate
-
pH 8.0, temperature not specified in the publication
27
-
malate
-
pH 9.5, temperature not specified in the publication
0.4
-
oxalate
-
10 mM
0.445
-
oxalate
-
1 mM
0.5
-
oxalate
-
intact enzyme
1.3
-
oxalate
-
cleaved enzyme
0.3
-
oxlate
-
-
-
3
-
pyruvate
-
competitive
6
-
pyruvate
-
using the FDH domain at varied ferrocyanide concentrations and a fixed L-lactate concentration pyruvate behaves as a mixed-type inhibitor toward ferrocyanide; using the FDH domain at varied L-lactate concentrations and a fixed ferrocyanide concentration pyruvate behaves as a mixed-type inhibitor toward L-lactate
9.7
-
pyruvate
-
in pre-steady-state kinetic studies of the FDH domain reduction and reoxidation, pyruvate acts as a competitive inhibitor of L-lactate
30
-
pyruvate
-
non-competitive
40
-
pyruvate
-
non-competitive
0.0073
-
zinc-substituted cytochrome c
-
pH 7.5, 25C, wild-type enzyme
-
SPECIFIC ACTIVITY [µmol/min/mg]
SPECIFIC ACTIVITY MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
0.055
-
-
(S)-lactate with cytochrome c as electron acceptor
0.4
-
-
+/- 0.02, was found in the cell-free extract
0.415
-
-
partially purified enzyme with 2,6-dichloroindophenol as electron acceptor
0.43
-
-
+/-0.05, cell-free extract
0.51
-
-
+/- 0.03, cell-free extract
0.66
-
-
+/- 0.06, the cell-free extract
0.72
-
-
6-dichloroindophenol as electron acceptor
3
-
-
cell-free extract from recombinant Hansenula polymorpha cell line expressing gene CYB2
3.2
-
-
recombinant enzyme in cells of Hansenula polymorpha
9
-
-
purified native enzyme, pH 7.5, 20C
895
-
-
purified enzyme
pH OPTIMUM
pH MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
5.5
-
-
before, after 10 days and 30 days rooting, substrate ferricyanide
6.3
7.5
-
assay at
7
7.5
-
assay at
7.2
8.4
-
-
7.5
-
-
before, after 30 days rooting, substrate ferricyanide
7.5
-
-
assay at
7.5
-
-
assay at
7.8
-
-
assay at
8
-
-
cleaved enzyme
8
-
-
assay at
9.5
-
-
before, after 10 days and 30 days rooting, substrate ferricyanide
pH RANGE
pH RANGE MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
5.2
-
-
almost no activity
TEMPERATURE OPTIMUM
TEMPERATURE OPTIMUM MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
20
24
-
assay at
20
-
-
assay at
25
30
-
assay at
30
-
-
assay at
pI VALUE
pI VALUE MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
SOURCE TISSUE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
SOURCE
-
optimization of culture conditions for recombinant cells of strain C-105 expressing gene CYB2 and showing increased FCb2 activity
Manually annotated by BRENDA team
-
two isozymes
Manually annotated by BRENDA team
PDB
SCOP
CATH
ORGANISM
Saccharomyces cerevisiae (strain ATCC 204508 / S288c)
Saccharomyces cerevisiae (strain ATCC 204508 / S288c)
Saccharomyces cerevisiae (strain ATCC 204508 / S288c)
Saccharomyces cerevisiae (strain ATCC 204508 / S288c)
Saccharomyces cerevisiae (strain ATCC 204508 / S288c)
Saccharomyces cerevisiae (strain ATCC 204508 / S288c)
Saccharomyces cerevisiae (strain ATCC 204508 / S288c)
Saccharomyces cerevisiae (strain ATCC 204508 / S288c)
Saccharomyces cerevisiae (strain ATCC 204508 / S288c)
Saccharomyces cerevisiae (strain ATCC 204508 / S288c)
Saccharomyces cerevisiae (strain ATCC 204508 / S288c)
Saccharomyces cerevisiae (strain ATCC 204508 / S288c)
MOLECULAR WEIGHT
MOLECULAR WEIGHT MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
10500
-
-
cytochrome domain, SDS-PAGE
56200
-
-
calculated from amino acid sequence, cleavage product from 64259 precursor
220000
-
-
gel filtration
220000
-
-
intact enzyme
230000
-
-
cleaved enzyme
235000
-
-
X-ray diffraction studies
SUBUNITS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
?
-
x * 47000, recombinantly expressed flavocytochrome b2 flavin-binding domain, SDS-PAGE
homotetramer
-
4 * 57500, X-ray crystallography
homotetramer
-
enzyme-substrate structure analysis during thr reaction cycle, overview
homotetramer
-
-
homotetramer
-
x-ray crystallography
homotetramer
-
each subunit of the soluble tetrameric enzyme consists of an N-terminal b5-like heme-binding domain and a C terminal flavodehydrogenase. The first 99 residues are folded around the heme, the next about 390 constitute the FMN-binding domain, and the last residues up to 511 make contacts with the other three subunits. Thus, the flavodehydrogenase domains constitute the core of the molecule, with a fourfold symmetry, while the heme domains lie at the periphery
homotetramer
Saccharomyces cerevisiae D273-10B/A1
-
-
-
tetramer
-
4 * 58600, calculated bases on amino acid sequence and heme extinction coefficient
tetramer
-
4 * 61000
tetramer
-
4 * 62750, SDS-PAGE
tetramer
-
4 * 34000-36000 + 4 * 21000-22000, cleaved enzyme; 4 * 57000
tetramer
-
4 * 60000
tetramer
-
4 * 57000
tetramer
-
4 * 57500, SDS-PAGE
Crystallization/COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
crystal structure of flavocytochrome b2 solved at 3.0 A resolution by the method of multiple isomorphous replacement with anomalous scattering
-
crystal structure of mutant H723Q bound with pyruvate is determined at 2.8 A
-
crystallization of flavin binding domain and intact enzyme, hanging drop vapor diffusion method
-
crystallization of Y143F mutant, vapor diffusion technique in presence of PEG 4000
-
Fcb2 free and in complex with sulfite, X-ray diffraction structure analysis at 2.3-2.6 A resolution
-
hanging drop vapor diffusion method, using 20% polyethylene glycol 4000, 0.1 M MES, pH 6.5, and 0.2 M MgCl2, at 18C
-
mutant L230A in complex with phenylglyoxalate, mutant A198G/L230A in complex with sulfite, mutant A198G/L230A in complex with pyruvate
-
used as essential purification step, crystallized as DNA complex
-
pH STABILITY
pH STABILITY MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
4
10
-
cytochrome domain stable
TEMPERATURE STABILITY
TEMPERATURE STABILITY MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
additional information
-
-
thermostable enzyme
GENERAL STABILITY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
intact enzyme unstable at low ionic strength
-
PMSF stabilizes, purification in absence of PMSF yields cleaved form of enzyme
-
STORAGE STABILITY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
-180C, long term storage
-
-70C in 0.1 M phosphate buffer, 1 mM EDTA, pH 7, 10 mM DL-lactate is added to the Fcb2 preparations to keep the enzyme in the reduced state
-
-70C, in 0.1 M phosphate buffer, 1 mM EDTA, pH 7, 10 mM DL-lactate is added to the Fcb2 preparations to keep the enzyme in the reduced state
-
-70C, purified enzymes are stored in 100 mM potassium phosphate, 1 mM EDTA, and 20 mM D,L-lactate at pH 7.5
-
-80C as saturated solution
-
-80C, 0.1 M phosphate buffer, 1 mM EDTA, 1 mM PMSF
-
4C, 70% saturated ammonium sulfate, under nitrogen, 2 months, 20% loss of activity of purified enzyme
-
4C, precipitate from 70% saturated ammonium sulfate, under nitrogen, stable for several weeks
-
under nitrogen, 1 month, no loss of activity
-
20C, 0.2 M phosphate buffer, pH 7.2, 1 mM EDTA, protected from light, 50% loss of activity within 10-20 days
-
4C, saturated ammonium sulfate, under nitrogen, protected from light, stabe
-
Purification/COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
native enzyme to homogeneity
-
cytochrome domain
-
partial purification of cytochrome b2 core
-
purification in presence and absence of PMSF results intact or cleaved enzyme
-
purification of different enzyme fragments
-
recombinantly expressed flavocytochrome b2 flavin-binding domain
-
wild type and mutant enzymes
-
flavin and heme domain from cloned Escherichia coli
-
Cloned/COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
gene CYB2, expression in the recipient strain Hansenula polymorpha C-105 impaired in glucose repression and devoid of catalase activity, under the control of the strong Hansenula polymorpha alcohol oxidase promoter in a plasmid for multicopy integration, optimization of culture conditions for recombinant cells with increased FCb2 activity
-
cytochrome domain in Escherichia coli
-
expressed in Saccharomyces cerevisiae YN1 strain (flavocytochrome b2 deletion mutant of strain XS560-1)
-
expression of flavocytochrome b2 flavin-binding domain in Escherichia coli
-
in Escherichia coli
-
in Escherichia coli AR120
-
in Escherichia coli JM 101
-
in Escherichia coli MM294
-
in Escherichia coli TG1
-
overexpression in Hansenula polymorpha leading to enhanced L-lactate-dependent respiration compared to the wild-type cells
-
recombinant wild-type Fcb2 and its recombinant FDH domain (FMN-binding domain) are expressed in Escherichia coli
-
recombinant wild-type Fcb2 and its recombinant flavin domain are expressed in Escherichia coli
-
recombinant wild-type Fcb2 and its recombinant FMN-binding domain (FDH domain) are expressed in Escherichia coli
-
flavin and heme domain expressed in Escherichia coli
-
in Escherichia coli JM 101
-
EXPRESSION
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
flavocytochrome b2 production is induced by the presence of oxygen and L-lactate
-
ENGINEERING
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
analysis
-
a L-lactate-selective microbial biosensor is developed using permeabilized cells of gene-engineered thermotolerant methylotrophic yeast Hansenula polymorpha, over-producing FCb2. The HpCYB2 gene, encoding FCb2, under the control of the strong Hansenula polymorpha alcohol oxidase promoter in the frame of a plasmid for multicopy integration is transformed to the recipient strain Hansenula polymorpha C-105 (gcr1 catX) impaired in glucose repression and devoid of catalase activity. The biosensor based on recombinant yeast cells exhibit a higher Km value (Km: 3.02 mM) and hence expanded linear range toward l-lactate as compared to a similar sensor based on the initial cells of Hansenula polymorpha C-105 (Km: 0.33 mM)
A198G
-
turnover reduced to 50%
A198G/L230A
-
double mutant, turnover reduced to less than 10%
A198G/L230A
-
double mutant enzyme shows significant activity towards L-mandelate
A198G/L230A
-
crystallization data
A67L
-
reaction proceeds slower than in wild type, no inhibition by monoclonal antibody inhibiting electron transfer via flavocytochrome b2
A67L
-
site-directed mutagenesis, comparison of the mutant kinetics in electron transfer from flavin to heme to the wild-type kinetics
A67Q
-
reaction proceeds slower than in wild type, no inhibition by monoclonal antibody inhibiting electron transfer via flavocytochrome b2
A67Q
-
site-directed mutagenesis, comparison of the mutant kinetics in electron transfer from flavin to heme to the wild-type kinetics
D72A
-
activity and inhibitory profile similar to wild type
D72A
-
site-directed mutagenesis, comparison of the mutant kinetics in electron transfer from flavin to heme to the wild-type kinetics
E63K
-
reaction proceeds slower than in wild type, no inhibition by monoclonal antibody inhibiting electron transfer via flavocytochrome b2
E63K
-
site-directed mutagenesis, comparison of the mutant kinetics in electron transfer from flavin to heme to the wild-type kinetics
E91K
-
mutation has no effect on the rate of cytochrome c reduction, no significantly different behavior with regard to inhibition by ferrocytochrome c
F39A
-
reaction proceeds slower than in wild type
F39A
-
site-directed mutagenesis, comparison of the mutant kinetics in electron transfer from flavin to heme to the wild-type kinetics
F52A
-
mutation has no effect on the rate of cytochrome c reduction
H373Q
-
His373 acts as an active site base during the oxidation of lactate to pyruvate. The decrease of 3500fold in the rate constant for reduction of the enzyme-bound FMN by lactate confirms this part of the reaction as that most affected by the mutation. Primary deuterium and solvent kinetic isotope affects for the mutant enzyme are significantly smaller than the wild-type values, establishing that bond cleavage steps are less rate-limiting in H373Q flavocytochrome b2 than in wild-type. Structure of the mutant enzyme with pyruvate bound, determined at 2.8 A, shows that the orientation of pyruvate in the active site is altered from that seen in the wild-type enzyme. Active site residues Arg289, Asp292, and Leu286 have altered positions in the mutant protein. The combination of an altered active site and the small kinetic isotope effects is consistent with the slowest step in turnover being a conformational change involving a conformation in which lactate is bound unproductively; kcat (1/sec) (substrate:lactate): 0.031 (intact protein), 0.057 (flavin domain only), Km (mM) (substrate: lactate): 0.65 (intact protein), 0.25 (flavin domain only)
H373Q
-
the mutation results in a 34 orders of magnitude decrease in kcat and a slight increase in L-lactate Km
K73A
-
activity and inhibitory profile similar to wild type
K73A
-
site-directed mutagenesis, comparison of the mutant kinetics in electron transfer from flavin to heme to the wild-type kinetics
L230A
-
turnover reduced to less than 10%
L230A
-
crystallization data
L230A
-
the mutant flavocytochrome b2 displays increased selectivity for (S)-2-hydroxyoctanoate over L-lactate by a factor of 40 (kcat/Km)
L230A/A198G
-
the double mutant enzyme has a 6fold greater catalytic efficiency with L-mandelate than with L-lactate
L65A
-
reaction proceeds slower than in wild type
N69K
-
reaction proceeds slower than in wild type, no inhibition by monoclonal antibody inhibiting electron transfer via flavocytochrome b2
N69K
-
site-directed mutagenesis, comparison of the mutant kinetics in electron transfer from flavin to heme to the wild-type kinetics
P44A
-
reaction proceeds slower than in wild type
P44A
-
site-directed mutagenesis, comparison of the mutant kinetics in electron transfer from flavin to heme to the wild-type kinetics
P64Q
-
less inhibition by monoclonal antibody inhibiting electron transfer via flavocytochrome b2
P64Q
-
site-directed mutagenesis, comparison of the mutant kinetics in electron transfer from flavin to heme to the wild-type kinetics
P64R
-
less inhibition by monoclonal antibody inhibiting electron transfer via flavocytochrome b2
P64R
-
site-directed mutagenesis, comparison of the mutant kinetics in electron transfer from flavin to heme to the wild-type kinetics
R289K
-
kcat (1/sec) (substrate: L-lactate): 8.6 (in 200 mM phosphate: 9.2, in 400 mM potassium acetate: 8.8, in 400 mM KCl: 9.2, in 400 mM KBr: 7.8), Km (mM) (substrate: L-lactate): 7.0 (in 200 mM phosphate: 8.7, in 400 mM potassium acetate: 9.2, in 400 mM KCl: 6.5, in 400 mM KBr: 5.8). Mutant is not sensitive for excess lactate concentration. In contrast to the wild-type enzyme high concentrations of acetate, phosphate, chloride and bromide show no influence on the mutant enzyme
R376K
-
mutant enzyme shows no activity
R38E
-
activity and inhibitory profile similar to wild type
V70M
-
less inhibition by monoclonal antibody inhibiting electron transfer via flavocytochromb2
V70M
-
site-directed mutagenesis, comparison of the mutant kinetics in electron transfer from flavin to heme to the wild-type kinetics
Y143F
-
turnover reduced to 15%
Y254del
-
deletion mutant
Y254F
-
significantly reduced activity of mutant enzyme
Y254F
-
only traces of activity
Y254F
-
increase in Km-value, 4fold decrease of vmax
Y254F
-
the mutant enzyme has a Vmax value some 28fold lower than that of the wild type enzyme and a slightly raised L-lactate Km value
Y254L
-
only traces of activity
Y254L
-
turnover reduced to less than 10%
Y74F
-
activity and inhibitory profile similar to wild type
Y74F
-
site-directed mutagenesis, comparison of the mutant kinetics in electron transfer from flavin to heme to the wild-type kinetics
L65A
-
site-directed mutagenesis, comparison of the mutant kinetics in electron transfer from flavin to heme to the wild-type kinetics
additional information
-
three different deletion mutants with deletion of 3, 6 and 9 amino acids from hinge region
additional information
-
the separately engineered flavodehydrogenase domain produces superoxide anion in its slow reaction with oxygen. This reaction apparently also takes place in the holoenzyme when oxygen is the sole electron acceptor, because the heme domain autoxidation is also slow. This is not unexpected in view of the heme domain mobility relative to the tetrameric flavodehydrogenase core. Reaction is so slow that it cannot compete with the normal electron flow in the presence of monoelectronic acceptors, such as ferricyanide and cytochrome c
additional information
-
introduction of a number of heme domain side chain substitutions in and around the interface to probe their effect on flavin to heme and cytochrome b2 electron transfer, overview
additional information
-
for detection of the enzyme activity, recombinant enzyme is immobilized by means of a dialysis membrane onto various types of electrode materials in order to investigate the possibility of electrochemically detecting L-lactate respiratio, overview
APPLICATION
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
analysis
-
sensitive and stable visualization of enzyme activity in cell-free extracts or during purification by by formation of precipitates of Berlin blue
analysis
-
development of a amperometric biosensor selective to L-lactate, bioanalytical properties are very fast response and high sensitivity and selectivity
environmental protection
-
the reductive pathway of the enzyme resulting in formation of less toxic Cr(III)-species is suggested to be the most important among possible mechanisms for chromate biodetoxification
synthesis
-
the enzyme is potentially important for bioanalytical technologies for highly selective assays of L-lactate in biological fluids and foods
environmental protection
Ogataea angusta C-105
-
the reductive pathway of the enzyme resulting in formation of less toxic Cr(III)-species is suggested to be the most important among possible mechanisms for chromate biodetoxification
-