Cloned (Comment) | Organism |
---|---|
expression in Escherichia coli | Clostridium autoethanogenum |
Protein Variants | Comment | Organism |
---|---|---|
G198D/S199 V/P201E/Y218A | mutant displays no detectable activity with NADPH as cofactor, but uses NADH with kcat/Km value of 12 per s and mM | Clostridium autoethanogenum |
additional information | variation of residue S199 does not yield any NADH-dependent enzymes | Clostridium autoethanogenum |
S199A | increase in catalytic efficiencies | Clostridium autoethanogenum |
S199C | decrease in catalytic efficiencies, with no significant changes in substrate preference | Clostridium autoethanogenum |
S199G | decrease in catalytic efficiencies, with no significant changes in substrate preference | Clostridium autoethanogenum |
S199R | increase in catalytic efficiencies | Clostridium autoethanogenum |
S199W | decrease in catalytic efficiencies, with no significant changes in substrate preference | Clostridium autoethanogenum |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.0017 | - |
NADPH | wild-type, pH 7.5, 25°C | Clostridium autoethanogenum | |
0.0056 | - |
NADPH | mutant S199A, pH 7.5, 25°C | Clostridium autoethanogenum | |
0.036 | - |
NADPH | mutant S199R, pH 7.5, 25°C | Clostridium autoethanogenum | |
0.8 | - |
2-butanone | mutant S199A, pH 7.5, 25°C | Clostridium autoethanogenum | |
0.9 | - |
acetone | mutant S199A, pH 7.5, 25°C | Clostridium autoethanogenum | |
1 | - |
acetone | wild-type, pH 7.5, 25°C | Clostridium autoethanogenum | |
1.1 | - |
acetone | mutant S199R, pH 7.5, 25°C | Clostridium autoethanogenum | |
1.2 | - |
acetone | mutant S199C, pH 7.5, 25°C | Clostridium autoethanogenum | |
1.5 | - |
2-butanone | mutant S199C, pH 7.5, 25°C | Clostridium autoethanogenum | |
1.6 | - |
acetone | mutant S199G, pH 7.5, 25°C | Clostridium autoethanogenum | |
1.8 | - |
2-butanone | wild-type, pH 7.5, 25°C | Clostridium autoethanogenum | |
1.9 | - |
2-butanone | mutant S199R, pH 7.5, 25°C | Clostridium autoethanogenum | |
2 | - |
acetone | mutant S199W, pH 7.5, 25°C | Clostridium autoethanogenum | |
2.4 | - |
2-butanone | mutant S199G, pH 7.5, 25°C | Clostridium autoethanogenum | |
3.6 | - |
2-butanone | mutant S199W, pH 7.5, 25°C | Clostridium autoethanogenum | |
34 | - |
(R)-acetoin | mutant S199R, pH 7.5, 25°C | Clostridium autoethanogenum | |
45 | - |
(R)-acetoin | mutant S199C, pH 7.5, 25°C | Clostridium autoethanogenum | |
98 | - |
(R)-acetoin | wild-type, pH 7.5, 25°C | Clostridium autoethanogenum | |
120 | - |
(R)-acetoin | mutant S199A, pH 7.5, 25°C | Clostridium autoethanogenum | |
190 | - |
(R)-acetoin | mutant S199W, pH 7.5, 25°C | Clostridium autoethanogenum | |
230 | - |
(R)-acetoin | mutant S199G, pH 7.5, 25°C | Clostridium autoethanogenum |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Clostridium autoethanogenum | F8TEL7 | - |
- |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
(R)-acetoin + NADPH + H+ | - |
Clostridium autoethanogenum | (2R,3R)-2,3-butanediol + NAD+ | - |
? | |
2-butanone + NADP+ | - |
Clostridium autoethanogenum | 2-butanol + NADPH + H+ | - |
? | |
acetone + NADPH + H+ | - |
Clostridium autoethanogenum | propan-2-ol + NADP+ | - |
? |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
2 | 8 | 2-butanone | mutant S199W, pH 7.5, 25°C | Clostridium autoethanogenum | |
3 | 6 | acetone | mutant S199C, pH 7.5, 25°C | Clostridium autoethanogenum | |
23 | - |
2-butanone | mutant S199C, pH 7.5, 25°C | Clostridium autoethanogenum | |
32 | - |
NADPH | mutant S199R, pH 7.5, 25°C | Clostridium autoethanogenum | |
37 | - |
2-butanone | wild-type, pH 7.5, 25°C | Clostridium autoethanogenum | |
40 | - |
2-butanone | mutant S199G, pH 7.5, 25°C | Clostridium autoethanogenum | |
41 | - |
(R)-acetoin | mutant S199C, pH 7.5, 25°C | Clostridium autoethanogenum | |
46 | - |
2-butanone | mutant S199A, pH 7.5, 25°C | Clostridium autoethanogenum | |
48 | - |
acetone | mutant S199W, pH 7.5, 25°C | Clostridium autoethanogenum | |
50 | - |
NADPH | mutant S199A, pH 7.5, 25°C | Clostridium autoethanogenum | |
50 | - |
2-butanone | mutant S199R, pH 7.5, 25°C | Clostridium autoethanogenum | |
55 | - |
acetone | wild-type, pH 7.5, 25°C | Clostridium autoethanogenum | |
60 | - |
acetone | mutant S199A, pH 7.5, 25°C | Clostridium autoethanogenum | |
62 | - |
acetone | mutant S199G, pH 7.5, 25°C | Clostridium autoethanogenum | |
66 | - |
(R)-acetoin | mutant S199W, pH 7.5, 25°C | Clostridium autoethanogenum | |
68 | - |
NADPH | wild-type, pH 7.5, 25°C | Clostridium autoethanogenum | |
89 | - |
acetone | mutant S199R, pH 7.5, 25°C | Clostridium autoethanogenum | |
95 | - |
(R)-acetoin | mutant S199R, pH 7.5, 25°C | Clostridium autoethanogenum | |
110 | - |
(R)-acetoin | mutant S199G, pH 7.5, 25°C | Clostridium autoethanogenum | |
150 | - |
(R)-acetoin | wild-type, pH 7.5, 25°C | Clostridium autoethanogenum | |
230 | - |
(R)-acetoin | mutant S199A, pH 7.5, 25°C | Clostridium autoethanogenum |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
NADH | cofoactor of mutant G198D/S199 V/P201E/Y218A | Clostridium autoethanogenum | |
NADPH | strictly dependent on NADPH | Clostridium autoethanogenum |
kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.35 | - |
(R)-acetoin | mutant S199W, pH 7.5, 25°C | Clostridium autoethanogenum | |
0.48 | - |
(R)-acetoin | mutant S199G, pH 7.5, 25°C | Clostridium autoethanogenum | |
0.9 | - |
(R)-acetoin | mutant S199C, pH 7.5, 25°C | Clostridium autoethanogenum | |
1.6 | - |
(R)-acetoin | wild-type, pH 7.5, 25°C | Clostridium autoethanogenum | |
1.9 | - |
(R)-acetoin | mutant S199A, pH 7.5, 25°C | Clostridium autoethanogenum | |
2.8 | - |
(R)-acetoin | mutant S199R, pH 7.5, 25°C | Clostridium autoethanogenum | |
7.8 | - |
2-butanone | mutant S199W, pH 7.5, 25°C | Clostridium autoethanogenum | |
15 | - |
2-butanone | mutant S199C, pH 7.5, 25°C | Clostridium autoethanogenum | |
17 | - |
2-butanone | mutant S199G, pH 7.5, 25°C | Clostridium autoethanogenum | |
21 | - |
2-butanone | wild-type, pH 7.5, 25°C | Clostridium autoethanogenum | |
24 | - |
acetone | mutant S199W, pH 7.5, 25°C | Clostridium autoethanogenum | |
26 | - |
2-butanone | mutant S199R, pH 7.5, 25°C | Clostridium autoethanogenum | |
30 | - |
acetone | mutant S199C, pH 7.5, 25°C | Clostridium autoethanogenum | |
39 | - |
acetone | mutant S199G, pH 7.5, 25°C | Clostridium autoethanogenum | |
55 | - |
acetone | wild-type, pH 7.5, 25°C | Clostridium autoethanogenum | |
61 | - |
2-butanone | mutant S199A, pH 7.5, 25°C | Clostridium autoethanogenum | |
67 | - |
acetone | mutant S199A, pH 7.5, 25°C | Clostridium autoethanogenum | |
81 | - |
acetone | mutant S199R, pH 7.5, 25°C | Clostridium autoethanogenum | |
890 | - |
NADPH | mutant S199R, pH 7.5, 25°C | Clostridium autoethanogenum | |
8900 | - |
NADPH | mutant S199A, pH 7.5, 25°C | Clostridium autoethanogenum | |
39000 | - |
NADPH | wild-type, pH 7.5, 25°C | Clostridium autoethanogenum |