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Literature summary for 1.1.2.3 extracted from

  • Gillet, N.; Ruiz-Pernia, J.J.; de la Lande, A.; Levy, B.; Lederer, F.; Demachy, I.; Moliner, V.
    QM/MM study of l-lactate oxidation by flavocytochrome b2 (2016), Phys. Chem. Chem. Phys., 18, 15609-15618 .
    View publication on PubMed

Protein Variants

Protein Variants Comment Organism
D282N site-directed mutagenesis, while the wild-type mutant has residue R289 in a distal or a proximal conformation, the mutant shows R289 only in a distal conformation Saccharomyces cerevisiae
additional information active site structures of wild-type and mutant enzymes, detailed overview Saccharomyces cerevisiae
Y254L site-directed mutagenesis, while the wild-type mutant has residue R289 in a distal or a proximal conformation, the mutant shows R289 only in a distal conformation Saccharomyces cerevisiae

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
additional information
-
additional information steady-state kinetics Saccharomyces cerevisiae

Localization

Localization Comment Organism GeneOntology No. Textmining
mitochondrial intermembrane space
-
Saccharomyces cerevisiae 5758
-

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
(S)-lactate + 2 ferricytochrome c Saccharomyces cerevisiae the first step of the catalytic cycle consists of the oxidation of L-lactate by the FMN prosthetic group. FMN is later reoxidized by transferring its electrons one by one to the ferric heme. The final electron acceptor is cytochrome c pyruvate + 2 ferrocytochrome c + 2 H+
-
?

Organism

Organism UniProt Comment Textmining
Saccharomyces cerevisiae P00175
-
-

Reaction

Reaction Comment Organism Reaction ID
(S)-lactate + 2 ferricytochrome c = pyruvate + 2 ferrocytochrome c + 2 H+ the hydride transfer mechanism involves the transfer of the lactate hydroxyl proton to H373 while the substrate aH atom is transferred as a hydride to the flavin mononucleotide (FMN) prosthetic group anchored in the active site Saccharomyces cerevisiae

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
(S)-lactate + 2 ferricytochrome c the first step of the catalytic cycle consists of the oxidation of L-lactate by the FMN prosthetic group. FMN is later reoxidized by transferring its electrons one by one to the ferric heme. The final electron acceptor is cytochrome c Saccharomyces cerevisiae pyruvate + 2 ferrocytochrome c + 2 H+
-
?
(S)-lactate + 2 flavocytochrome b2
-
Saccharomyces cerevisiae pyruvate + 2 ferrocytochrome c + 2 H+
-
?

Subunits

Subunits Comment Organism
homotetramer
-
Saccharomyces cerevisiae

Synonyms

Synonyms Comment Organism
Fcb2
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Saccharomyces cerevisiae
L-lactate cytochrome c oxidoreductase
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Saccharomyces cerevisiae

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
30
-
assay at Saccharomyces cerevisiae

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
0.51
-
(S)-lactate mutant Y254L with R289 distal, pH 7.0, 30°C Saccharomyces cerevisiae
3.9
-
(S)-lactate mutant D282N with R289 distal, pH 7.0, 30°C Saccharomyces cerevisiae
270
-
(S)-lactate wild-type enzyme with R289 distal, pH 7.0, 30°C Saccharomyces cerevisiae

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
7
-
assay at Saccharomyces cerevisiae

Cofactor

Cofactor Comment Organism Structure
ferricytochrome c
-
Saccharomyces cerevisiae
flavocytochrome b2
-
Saccharomyces cerevisiae
FMN each of the four subunits contains a flavin mononucleotide prosthetic group Saccharomyces cerevisiae
heme each of the four subunits contains a beta-type heme Saccharomyces cerevisiae
additional information FMN and heme groups are localized in two different domains Saccharomyces cerevisiae

General Information

General Information Comment Organism
additional information molecular dynamics simulations using a hybrid quantum mechanics/molecular mechanics (QM/MM) scheme to study the mechanism of L-lactate oxidation by flavocytochrome b2. Simulation results highlight the influence of the environment on the catalytic mechanism by describing a step-wise process in the wild-type enzyme with R289 in a distal position and a concerted mechanism for the other systems. Structure analysis of pyruvate in the Fcb2 active site pocket with R289 in the distal conformation. Residue Y254 plays a role in the catalytic process by stabilizing the product of the first proton transfer from substrate to H373, while residue D282 is expected to stabilize the imidazolium ion in transition and product states by electrostatic interactions and hydrogen bonding with the Hdelta of H373. Active site structures of wild-type and mutant enzymes, detailed overview Saccharomyces cerevisiae