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Literature summary extracted from
- Potentiometric and further kinetic characterization of the flavin-binding domain of Saccharomyces cerevisiae flavocytochrome b2. Inhibition by anions binding in the active site (2007), Biochemistry, 46, 4661-4670.
Cloned(Commentary)
| EC Number | Cloned (Comment) | Organism |
|---|---|---|
| 1.1.2.3 | recombinant wild-type Fcb2 and its recombinant FMN-binding domain (FDH domain) are expressed in Escherichia coli | Saccharomyces cerevisiae |
Protein Variants
| EC Number | Protein Variants | Comment | Organism |
|---|---|---|---|
| 1.1.2.3 | 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 | Saccharomyces cerevisiae |
Inhibitors
| EC Number | Inhibitors | Comment | Organism | Structure |
|---|---|---|---|---|
| 1.1.2.3 | acetate | - |
Saccharomyces cerevisiae |  |
| 1.1.2.3 | bromide | - |
Saccharomyces cerevisiae | |
| 1.1.2.3 | chloride | - |
Saccharomyces cerevisiae | |
| 1.1.2.3 | additional information | enzyme is inhibited by high substrate concentrations | Saccharomyces cerevisiae | |
| 1.1.2.3 | phosphate | - |
Saccharomyces cerevisiae | |
| 1.1.2.3 | pyruvate | - |
Saccharomyces cerevisiae | |
KM Value [mM]
| EC Number | KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|---|
| 1.1.2.3 | 0.66 | - |
L-lactate | FDH domain, reaction conditions: 13 mM ferricyanide, 100 mM phosphate buffer, 1 mM EDTA, pH 7, 30°C | Saccharomyces cerevisiae | |
| 1.1.2.3 | 0.66 | - |
L-lactate | FDH domain, reaction conditions: 13 mM ferricyanide, 100 mM phosphate buffer, 1 mM EDTA, pH 7.5, 25°C | Saccharomyces cerevisiae | |
| 1.1.2.3 | 0.86 | - |
L-lactate | FDH domain, reaction conditions: 13 mM ferricyanide, 10 mM Tris/HCl buffer, 0.1 M NaCl, pH 7.5, 25°C | Saccharomyces cerevisiae | |
| 1.1.2.3 | 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, 5°C, in the absence of ferrocyanide | Saccharomyces cerevisiae | |
| 1.1.2.3 | 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, 5°C, 1.5 mM ferricyanide and variable L-lactate concentration | Saccharomyces cerevisiae | |
| 1.1.2.3 | 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, 5°C, 1.5 mM ferricyanide and variable L-lactate concentration | Saccharomyces cerevisiae | |
| 1.1.2.3 | 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, 5°C, in the absence of ferrocyanide | Saccharomyces cerevisiae | |
| 1.1.2.3 | 1.18 | - |
L-lactate | FDH domain, reaction conditions: 13 mM ferricyanide, 100 mM phosphate buffer, 1 mM EDTA, pH 7, 5°C | Saccharomyces cerevisiae | |
| 1.1.2.3 | 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, 5°C, 1.5 mM ferricyanide and variable L-lactate concentration | Saccharomyces cerevisiae | |
| 1.1.2.3 | 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, 5°C, constant 10 mM ferricyanide and variable L-lactate concentration | Saccharomyces cerevisiae | |
| 1.1.2.3 | 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, 5°C, 1.5 mM ferricyanide and variable L-lactate concentration | Saccharomyces cerevisiae | |
| 1.1.2.3 | 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, 5°C, constant 10 mM ferricyanide and variable L-lactate concentration | Saccharomyces cerevisiae | |
| 1.1.2.3 | 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, 5°C, constant 10 mM ferricyanide and variable L-lactate concentration | Saccharomyces cerevisiae | |
| 1.1.2.3 | 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, 5°C, constant 10 mM ferricyanide and variable L-lactate concentration | Saccharomyces cerevisiae | |
| 1.1.2.3 | 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, 5°C, 2 mM ferricyanide and variable L-lactate concentration | Saccharomyces cerevisiae | |
| 1.1.2.3 | 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, 5°C, 2 mM ferricyanide and variable L-lactate concentration | Saccharomyces cerevisiae | |
| 1.1.2.3 | 7 | - |
L-lactate | mutant R289K steady-state kinetic. Reaction conditions: 100 mM phosphate buffer, 1 mM EDTA, pH 7, 5°C, 2 mM ferricyanide and variable L-lactate concentration | Saccharomyces cerevisiae | |
| 1.1.2.3 | 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, 5°C, 2 mM ferricyanide and variable L-lactate concentration | Saccharomyces cerevisiae | |
| 1.1.2.3 | 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, 5°C, 2 mM ferricyanide and variable L-lactate concentration | Saccharomyces cerevisiae | |
Organism
| EC Number | Organism | UniProt | Comment | Textmining |
|---|---|---|---|---|
| 1.1.2.3 | Saccharomyces cerevisiae | - |
- |
- |
Storage Stability
| EC Number | Storage Stability | Organism |
|---|---|---|
| 1.1.2.3 | -70°C 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 | Saccharomyces cerevisiae |
Substrates and Products (Substrate)
| EC Number | Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
|---|---|---|---|---|---|---|---|
| 1.1.2.3 | L-lactate + ferricyanide | - |
Saccharomyces cerevisiae | pyruvate + ferrocyanide + H+ | - |
? | |
Synonyms
| EC Number | Synonyms | Comment | Organism |
|---|---|---|---|
| 1.1.2.3 | cytochrome c oxido reductase | - |
Saccharomyces cerevisiae |
| 1.1.2.3 | flavocytochrome b2 | - |
Saccharomyces cerevisiae |
Turnover Number [1/s]
| EC Number | Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|---|
| 1.1.2.3 | 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, 5°C, 2 mM ferricyanide and variable L-lactate concentration | Saccharomyces cerevisiae | |
| 1.1.2.3 | 8.6 | - |
L-lactate | mutant R289K steady-state kinetic. Reaction conditions: 100 mM phosphate buffer, 1 mM EDTA, pH 7, 5°C, 2 mM ferricyanide and variable L-lactate concentration | Saccharomyces cerevisiae | |
| 1.1.2.3 | 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, 5°C, 2 mM ferricyanide and variable L-lactate concentration | Saccharomyces cerevisiae | |
| 1.1.2.3 | 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, 5°C, 2 mM ferricyanide and variable L-lactate concentration | Saccharomyces cerevisiae | |
| 1.1.2.3 | 9.2 | - |
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, 5°C, 2 mM ferricyanide and variable L-lactate concentration | Saccharomyces cerevisiae | |
| 1.1.2.3 | 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, 5°C, constant 10 mM ferricyanide and variable L-lactate concentration | Saccharomyces cerevisiae | |
| 1.1.2.3 | 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, 5°C, constant 10 mM ferricyanide and variable L-lactate concentration | Saccharomyces cerevisiae | |
| 1.1.2.3 | 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, 5°C, 1.5 mM ferricyanide and variable L-lactate concentration | Saccharomyces cerevisiae | |
| 1.1.2.3 | 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, 5°C, 1.5 mM ferricyanide and variable L-lactate concentration | Saccharomyces cerevisiae | |
| 1.1.2.3 | 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, 5°C, 1.5 mM ferricyanide and variable L-lactate concentration | Saccharomyces cerevisiae | |
| 1.1.2.3 | 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, 5°C, constant 10 mM ferricyanide and variable L-lactate concentration | Saccharomyces cerevisiae | |
| 1.1.2.3 | 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, 5°C, constant 10 mM ferricyanide and variable L-lactate concentration | Saccharomyces cerevisiae | |
| 1.1.2.3 | 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, 5°C, 1.5 mM ferricyanide and variable L-lactate concentration | Saccharomyces cerevisiae | |
| 1.1.2.3 | 117 | - |
L-lactate | holo-enzyme, reaction conditions: 1 or 2 mM ferricyanide, 100 mM phosphate buffer, 1mM EDTA, pH 7, 5°C | Saccharomyces cerevisiae | |
| 1.1.2.3 | 133 | - |
L-lactate | FDH domain, reaction conditions: 13 mM ferricyanide, 100 mM phosphate buffer, 1 mM EDTA, pH 7, 5°C | Saccharomyces cerevisiae | |
| 1.1.2.3 | 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, 5°C, in the absence of ferrocyanide | Saccharomyces cerevisiae | |
| 1.1.2.3 | 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, 5°C, in the absence of ferrocyanide | Saccharomyces cerevisiae | |
| 1.1.2.3 | 214 | - |
L-lactate | FDH domain, reaction conditions: 13 mM ferricyanide, 10 mM Tris/HCl buffer, 0.1 M NaCl, pH 7.5, 25°C | Saccharomyces cerevisiae | |
| 1.1.2.3 | 240 | - |
L-lactate | FDH domain, reaction conditions: 13 mM ferricyanide, 100 mM phosphate buffer, 1 mM EDTA, pH 7.5, 25°C | Saccharomyces cerevisiae | |
| 1.1.2.3 | 259 | - |
L-lactate | FDH domain, reaction conditions: 13 mM ferricyanide, 100 mM phosphate buffer, 1 mM EDTA, pH 7, 30°C | Saccharomyces cerevisiae | |
Cofactor
| EC Number | Cofactor | Comment | Organism | Structure |
|---|---|---|---|---|
| 1.1.2.3 | FMN | - |
Saccharomyces cerevisiae | |
Ki Value [mM]
| EC Number | Ki Value [mM] | Ki Value maximum [mM] | Inhibitor | Comment | Organism | Structure |
|---|---|---|---|---|---|---|
| 1.1.2.3 | 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 | Saccharomyces cerevisiae | |
| 1.1.2.3 | 6 | - |
pyruvate | using the FDH domain at varied L-lactate concentrations and a fixed ferrocyanide concentration pyruvate behaves as a mixed-type inhibitor toward L-lactate | Saccharomyces cerevisiae | |
| 1.1.2.3 | 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 | Saccharomyces cerevisiae | |
| 1.1.2.3 | 170 | - |
acetate | in pre-steady-state kinetic studies of the FDH domain reduction and reoxidation, acetate acts as a competitive inhibitor of L-lactate | Saccharomyces cerevisiae | |
| 1.1.2.3 | 450 | - |
chloride | in pre-steady-state kinetic studies of the FDH domain reduction and reoxidation, chloride acts as a competitive inhibitor of L-lactate | Saccharomyces cerevisiae | |
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