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Literature summary for 1.8.1.4 extracted from

  • Hakansson, A.P.; Smith, A.W.
    Enzymatic characterization of dihydrolipoamide dehydrogenase from Streptococcus pneumoniae harboring its own substrate (2007), J. Biol. Chem., 282, 29521-29530.
    View publication on PubMed

Application

Application Comment Organism
additional information the lipoyl protein domain (but not lipoic acid alone) plays a regulatory role in the enzymatic characteristics of pneumococcal DLDH Streptococcus pneumoniae

Cloned(Commentary)

Cloned (Comment) Organism
expressed in Escherichia coli M15 cells Streptococcus pneumoniae

Protein Variants

Protein Variants Comment Organism
K43A unable to express the mutant protein Streptococcus pneumoniae
K43R expresses well, but still has lipoic acid attached Streptococcus pneumoniae
additional information protein expressed without lipoic acid is indistinguishable from the wild-type protein. The protein without a lipoyl protein domain has a 2-3fold higher turnover number, a lower Ki for NADH, and a higher Ki for lipoamide compared with the other two enzymes Streptococcus pneumoniae

Inhibitors

Inhibitors Comment Organism Structure
Lipoamide
-
Streptococcus pneumoniae
NADH
-
Streptococcus pneumoniae
potassium phosphate when purified DLDH is eluted directly into potassium phosphate buffer, the enzymatic activity rapidly decreases Streptococcus pneumoniae

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.04
-
dihydrolipoamide recombinant DLDH expressed without without a lipoyl protein domain Streptococcus pneumoniae
0.056
-
dihydrolipoamide recombinant DLDH Streptococcus pneumoniae
0.064
-
dihydrolipoamide recombinant DLDH expressed without lipoic acid Streptococcus pneumoniae
0.82
-
NAD+ recombinant DLDH expressed without without a lipoyl protein domain Streptococcus pneumoniae
0.985
-
NAD+ recombinant DLDH expressed without lipoic acid Streptococcus pneumoniae
1.037
-
NAD+ recombinant DLDH Streptococcus pneumoniae

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
61600
-
full-length recombinant DLDH, sequence analysis Streptococcus pneumoniae

Organism

Organism UniProt Comment Textmining
Streptococcus pneumoniae
-
-
-

Purification (Commentary)

Purification (Comment) Organism
gel filtration, at least 90% pure Streptococcus pneumoniae

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
dihydrolipoamide + NAD+
-
Streptococcus pneumoniae lipoamide + NADH
-
r
lipoamide + NADH
-
Streptococcus pneumoniae dihydrolipoamide + NAD+
-
r
nitro blue tetrazolium + NADH
-
Streptococcus pneumoniae ? + NAD+
-
?

Synonyms

Synonyms Comment Organism
dihydrolipoamide dehydrogenase
-
Streptococcus pneumoniae
DLDH
-
Streptococcus pneumoniae

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
160
-
NAD+ recombinant DLDH Streptococcus pneumoniae
223
-
NAD+ recombinant DLDH expressed without lipoic acid Streptococcus pneumoniae
506
-
NAD+ recombinant DLDH expressed without without a lipoyl protein domain Streptococcus pneumoniae

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
8.3
-
in the reverse reaction Streptococcus pneumoniae
8.5
-
in the forward reaction Streptococcus pneumoniae

Cofactor

Cofactor Comment Organism Structure
FAD
-
Streptococcus pneumoniae
additional information unable to use NADP+ and NADPH Streptococcus pneumoniae
NAD+
-
Streptococcus pneumoniae
NADH
-
Streptococcus pneumoniae

Ki Value [mM]

Ki Value [mM] Ki Value maximum [mM] Inhibitor Comment Organism Structure
0.0044
-
NADH recombinant DLDH expressed without without a lipoyl protein domain, in the presence of 1.5 mM dihydrolipoamide Streptococcus pneumoniae
0.0107
-
NADH recombinant DLDH expressed without lipoic acid, in the presence of 1.5 mM dihydrolipoamide Streptococcus pneumoniae
0.0149
-
NADH recombinant DLDH, in the presence of 1.5 mM dihydrolipoamide Streptococcus pneumoniae
0.0351
-
NADH recombinant DLDH expressed without without a lipoyl protein domain, in the presence of 2 mM NAD+ Streptococcus pneumoniae
0.0488
-
Lipoamide recombinant DLDH expressed without lipoic acid, in the presence of 4 mM NAD+ Streptococcus pneumoniae
0.0566
-
Lipoamide recombinant DLDH, in the presence of 4 mM NAD+ Streptococcus pneumoniae
0.0722
-
NADH recombinant DLDH, in the presence of 2 mM NAD+ Streptococcus pneumoniae
0.084
-
NADH recombinant DLDH expressed without lipoic acid, in the presence of 2 mM NAD+ Streptococcus pneumoniae
0.1899
-
Lipoamide recombinant DLDH expressed without lipoic acid, in the presence of 1.5 mM dihydrolipoamide Streptococcus pneumoniae
0.1953
-
Lipoamide recombinant DLDH, in the presence of 1.5 mM dihydrolipoamide Streptococcus pneumoniae
0.349
-
Lipoamide recombinant DLDH expressed without without a lipoyl protein domain, in the presence of 4 mM NAD+ Streptococcus pneumoniae
0.8793
-
Lipoamide recombinant DLDH expressed without without a lipoyl protein domain, in the presence of 1.5 mM dihydrolipoamide Streptococcus pneumoniae