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Literature summary for 1.1.3.2 extracted from

  • Stoisser, T.; Brunsteiner, M.; Wilson, D.K.; Nidetzky, B.
    Conformational flexibility related to enzyme activity evidence for a dynamic active-site gatekeeper function of Tyr(215) in Aerococcus viridans lactate oxidase (2016), Sci. Rep., 6, 27892 .
    View publication on PubMedView publication on EuropePMC
Search for more literature for 1.1.3.2

Crystallization (Commentary)

Crystallization (Comment) Organism
mutant Y215F in complex with pyruvate, to 2.6 A resolution. The hydrogen bond between the phenolic hydroxyl and the keto oxygen in pyruvate is replaced with a potentially stronger hydrophobic interaction between the phenylalanine and the methyl group of pyruvate. Residues 200 through 215 or 216 appear to be disordered Aerococcus viridans

Protein Variants

Protein Variants Comment Organism
Y215F mutation in binding pocket, mutant shows slowed flavin reduction and oxidation by up to 33-fold. Pyruvate release is also decelerated and is the slowest step overall Aerococcus viridans
Y215H Y215F mutation in binding pocket, mutant shows slowed flavin reduction and oxidation by up to 33-fold. Pyruvate release is also decelerated Aerococcus viridans

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.024
-
 O2 mutant Y215F, pH 6.5, 37°C Aerococcus viridans
0.051
-
 O2 wild-type, pH 6.5, 37°C Aerococcus viridans
0.062
-
 O2 mutant Y215H, pH 6.5, 37°C Aerococcus viridans
0.13
-
 (S)-lactate mutant Y215F, pH 6.5, 37°C Aerococcus viridans
0.5
-
 (S)-lactate wild-type, pH 6.5, 37°C Aerococcus viridans
0.52
-
 (S)-lactate mutant Y215H, pH 6.5, 37°C Aerococcus viridans

Organism

Organism UniProt Comment Textmining
Aerococcus viridans Q44467
-
-

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
(S)-lactate + O2
-
 Aerococcus viridans pyruvate + H2O2
-
 ?

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
5
-
 (S)-lactate mutant Y215H, pH 6.5, 37°C Aerococcus viridans
22
-
 (S)-lactate mutant Y215F, pH 6.5, 37°C Aerococcus viridans
88
-
 (S)-lactate wild-type, pH 6.5, 37°C Aerococcus viridans

kcat/KM [mM/s]

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
9.61
-
 (S)-lactate mutant Y215H, pH 6.5, 37°C Aerococcus viridans
169
-
 (S)-lactate mutant Y215F, pH 6.5, 37°C Aerococcus viridans
176
-
 (S)-lactate wild-type, pH 6.5, 37°C Aerococcus viridans