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Literature summary for 1.8.1.4 extracted from

  • Kim, Y.; Ingram, L.O.; Shanmugam, K.T.
    Dihydrolipoamide dehydrogenase mutation alters the NADH sensitivity of pyruvate dehydrogenase complex of Escherichia coli K-12 (2008), J. Bacteriol., 190, 3851-3858.
    View publication on PubMedView publication on EuropePMC
Search for more literature for 1.8.1.4

Application

Application Comment Organism
additional information a mutation in LPD leads to a pyruvate dehydrogenase complex that is less sensitive to inhibition by NADH, allowing the enzyme to function in an anaerobic culture, which changes the fermentation profile of the mutant. Presence and functional activity of such an NADH-insensitive pyruvate dehydrogenase may have significant unexplored physiological and biotechnological applications Escherichia coli K-12

Cloned(Commentary)

Cloned (Comment) Organism
expressed in Escherichia coli M109(lambdaDE3) Escherichia coli K-12

Protein Variants

Protein Variants Comment Organism
E354K is significantly less sensitive to NADH inhibition than the native LPD Escherichia coli K-12
H322Y upon purification LPD loses activity and associated FAD at the gel filtration step Escherichia coli K-12

Inhibitors

Inhibitors Comment Organism Structure
FAD
-
 Escherichia coli K-12
NADH
-
 Escherichia coli K-12

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.07
-
 NAD+ native LPD Escherichia coli K-12
0.08
-
 NAD+ mutant E354K Escherichia coli K-12
0.11
-
 NAD+ mutant H322Y Escherichia coli K-12
0.3
-
 pyruvate mutant E354K Escherichia coli K-12
0.4
-
 pyruvate native LPD Escherichia coli K-12
0.48
-
 pyruvate mutant H322Y Escherichia coli K-12

Localization

Localization Comment Organism GeneOntology No. Textmining

Organism

Organism UniProt Comment Textmining
Escherichia coli K-12
-
 strains AH242 SE2378 and SE2382
-

Purification (Commentary)

Purification (Comment) Organism
gel filtration Escherichia coli K-12

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
lipoamide + NADH
-
 Escherichia coli K-12 dihydrolipoamide + NAD+
-
 r
pyruvate + NADH
-
 Escherichia coli K-12 ?
-
 ?

Synonyms

Synonyms Comment Organism
dihydrolipoamide dehydrogenase
-
 Escherichia coli K-12
LPD
-
 Escherichia coli K-12

Cofactor

Cofactor Comment Organism Structure
NAD+
-
 Escherichia coli K-12
NADH
-
 Escherichia coli K-12

Ki Value [mM]

Ki Value [mM] Ki Value maximum [mM] Inhibitor Comment Organism Structure
0.001
-
 NADH native pyruvate dehydrogenase complex Escherichia coli K-12
0.0096
-
 NADH mutant H322Y Escherichia coli K-12
0.01
-
 NADH mutant E354K Escherichia coli K-12