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Literature summary for 1.17.1.10 extracted from

  • Hoelsch, K.; Suehrer, I.; Heusel, M.; Weuster-Botz, D.
    Engineering of formate dehydrogenase: synergistic effect of mutations affecting cofactor specificity and chemical stability (2013), Appl. Microbiol. Biotechnol., 97, 2473-2481.
    View publication on PubMed

Cloned(Commentary)

Cloned (Comment) Organism
expressed in Escherichia coli BL21(DE3) cells Mycolicibacterium vaccae

Protein Variants

Protein Variants Comment Organism
A198G/D221Q the mutant has the highest catalytic efficiency with NADP+ Mycolicibacterium vaccae
C145S/A198G/D221Q/C255V the mutant has high catalytic efficiency with NADP+ Mycolicibacterium vaccae
C145S/D221A/C255V the mutant has high catalytic efficiency with NADP+ Mycolicibacterium vaccae
C145S/D221Q/C255V the mutant shows the highest specific activity for a NADP+-accepting formate dehydrogenase Mycolicibacterium vaccae
D221A the mutant has high catalytic efficiency with NADP+ Mycolicibacterium vaccae
D221G the mutant has high catalytic efficiency with NADP+ Mycolicibacterium vaccae
D221Q the mutant has high catalytic efficiency with NADP+ Mycolicibacterium vaccae
D221S the mutant has high catalytic efficiency with NADP+ Mycolicibacterium vaccae

Inhibitors

Inhibitors Comment Organism Structure
ethyl 4-chloroacetoacetate
-
Mycolicibacterium vaccae

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.084
-
NADP+ mutant C145S/D221Q/C255V, at pH 7.0 and 22°C Mycolicibacterium vaccae
0.147
-
NADP+ mutant C145S/A198G/D221Q/C255V, at pH 7.0 and 22°C Mycolicibacterium vaccae
0.24
-
NADP+ mutant D221G/C255V, at pH 7.0 and 22°C Mycolicibacterium vaccae
0.27
-
NADP+ mutant A198G/D221Q, at pH 7.0 and 22°C Mycolicibacterium vaccae
0.31
-
NADP+ mutant D221G, at pH 7.0 and 22°C Mycolicibacterium vaccae
0.37
-
NADP+ mutant D221A, at pH 7.0 and 22°C Mycolicibacterium vaccae
0.39
-
NADP+ mutant D221Q, at pH 7.0 and 22°C Mycolicibacterium vaccae
0.43
-
NADP+ mutant D221S, at pH 7.0 and 22°C Mycolicibacterium vaccae
0.57
-
NADP+ mutant C145S/D221G/C255V, at pH 7.0 and 22°C Mycolicibacterium vaccae
0.92
-
NADP+ mutant C145S/D221A/C255V, at pH 7.0 and 22°C Mycolicibacterium vaccae
2 3 formate mutant D221A, at pH 7.0 and 22°C Mycolicibacterium vaccae
40
-
formate mutant D221G, at pH 7.0 and 22°C Mycolicibacterium vaccae
41
-
formate mutant D221S, at pH 7.0 and 22°C Mycolicibacterium vaccae
46
-
formate mutant D221Q, at pH 7.0 and 22°C Mycolicibacterium vaccae
62
-
formate mutant D221G/C255V, at pH 7.0 and 22°C Mycolicibacterium vaccae
69
-
formate mutant C145S/D221G/C255V, at pH 7.0 and 22°C Mycolicibacterium vaccae
127
-
formate mutant A198G/D221Q, at pH 7.0 and 22°C Mycolicibacterium vaccae

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
formate + NADP+ Mycolicibacterium vaccae
-
CO2 + NADPH
-
?
formate + NADP+ Mycolicibacterium vaccae N10
-
CO2 + NADPH
-
?

Organism

Organism UniProt Comment Textmining
Mycolicibacterium vaccae
-
-
-
Mycolicibacterium vaccae N10
-
-
-

Purification (Commentary)

Purification (Comment) Organism
HisTrap column chromatography Mycolicibacterium vaccae

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
formate + NADP+
-
Mycolicibacterium vaccae CO2 + NADPH
-
?
formate + NADP+
-
Mycolicibacterium vaccae N10 CO2 + NADPH
-
?

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
0.52
-
NADP+ mutant D221G, at pH 7.0 and 22°C Mycolicibacterium vaccae
0.55
-
NADP+ mutant D221S, at pH 7.0 and 22°C Mycolicibacterium vaccae
0.62
-
NADP+ mutant D221G/C255V, at pH 7.0 and 22°C Mycolicibacterium vaccae
0.79
-
NADP+ mutant A198G/D221Q, at pH 7.0 and 22°C Mycolicibacterium vaccae
1.04
-
NADP+ mutant D221Q, at pH 7.0 and 22°C Mycolicibacterium vaccae
1.05
-
NADP+ mutant D221A, at pH 7.0 and 22°C Mycolicibacterium vaccae
1.5
-
NADP+ mutant C145S/D221Q/C255V, at pH 7.0 and 22°C Mycolicibacterium vaccae
1.64
-
NADP+ mutant C145S/D221G/C255V, at pH 7.0 and 22°C Mycolicibacterium vaccae
3.08
-
NADP+ mutant C145S/A198G/D221Q/C255V, at pH 7.0 and 22°C Mycolicibacterium vaccae
7.89
-
NADP+ mutant C145S/D221A/C255V, at pH 7.0 and 22°C Mycolicibacterium vaccae

Cofactor

Cofactor Comment Organism Structure
NADP+
-
Mycolicibacterium vaccae

kcat/KM [mM/s]

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
1.29
-
NADP+ mutant D221S, at pH 7.0 and 22°C Mycolicibacterium vaccae
1.69
-
NADP+ mutant D221G, at pH 7.0 and 22°C Mycolicibacterium vaccae
2.58
-
NADP+ mutant D221G/C255V, at pH 7.0 and 22°C Mycolicibacterium vaccae
2.67
-
NADP+ mutant D221Q, at pH 7.0 and 22°C Mycolicibacterium vaccae
2.83
-
NADP+ mutant D221A, at pH 7.0 and 22°C Mycolicibacterium vaccae
2.88
-
NADP+ mutant C145S/D221G/C255V, at pH 7.0 and 22°C Mycolicibacterium vaccae
2.94
-
NADP+ mutant A198G/D221Q, at pH 7.0 and 22°C Mycolicibacterium vaccae
8.58
-
NADP+ mutant C145S/D221A/C255V, at pH 7.0 and 22°C Mycolicibacterium vaccae
17.9
-
NADP+ mutant C145S/D221Q/C255V, at pH 7.0 and 22°C Mycolicibacterium vaccae
21
-
NADP+ mutant C145S/A198G/D221Q/C255V, at pH 7.0 and 22°C Mycolicibacterium vaccae