Cloned (Comment) | Organism |
---|---|
expression in Escherichia coli | Xanthobacter autotrophicus |
Protein Variants | Comment | Organism |
---|---|---|
K160A | inactive | Xanthobacter autotrophicus |
K214A | substantially reduced catalytic efficiencies with the natural substrates in both the oxidative and reductive direction | Xanthobacter autotrophicus |
R211A | substantially reduced catalytic efficiencies with the natural substrates in both the oxidative and reductive direction | Xanthobacter autotrophicus |
S143A | residue acts as H-bond donor to substrate and in charge stabilization of the transition state, large increase in Km value | Xanthobacter autotrophicus |
Y156A | mutation in general acid/base, large increase in Km value | Xanthobacter autotrophicus |
Y156F | inactive | Xanthobacter autotrophicus |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
2-(2-methyl-2-hydroxypropylthio)ethanesulfonate | competitive; competitive | Xanthobacter autotrophicus |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.031 | - |
2-(S)-hydroxypropyl-CoM | wild-type, pH 7.5, 30°C | Xanthobacter autotrophicus | |
0.068 | - |
2-oxopropyl-CoM | pH 7.5, 30°C | Xanthobacter autotrophicus | |
0.096 | - |
2-(R)-hydroxypropyl-CoM | pH 7.5, 30°C | Xanthobacter autotrophicus | |
0.22 | - |
2-(S)-hydroxypropyl-CoM | pH 7.5, 30°C | Xanthobacter autotrophicus | |
0.27 | - |
2-oxopropyl-CoM | wild-type, pH 7.5, 30°C | Xanthobacter autotrophicus | |
0.97 | - |
2-(2-hydroxyethylthio)ethanesulfonate | wild-type, pH 7.5, 30°C | Xanthobacter autotrophicus | |
1.6 | - |
2-(S)-hydroxypropyl-CoM | mutant R211A, pH 7.5, 30°C | Xanthobacter autotrophicus | |
1.8 | - |
2-(S)-hydroxypropyl-CoM | mutant Y156A, pH 7.5, 30°C | Xanthobacter autotrophicus | |
2.3 | - |
2-(S)-hydroxypropyl-CoM | mutant K214A, pH 7.5, 30°C | Xanthobacter autotrophicus | |
9.1 | - |
2-(R)-hydroxypropyl-CoM | wild-type, pH 7.5, 30°C | Xanthobacter autotrophicus | |
9.5 | - |
2-(S)-hydroxypropyl-CoM | mutant S143A, pH 7.5, 30°C | Xanthobacter autotrophicus | |
11 | - |
2-oxopropyl-CoM | mutant R211A, pH 7.5, 30°C | Xanthobacter autotrophicus | |
12 | - |
2-oxopropyl-CoM | mutant K214A, pH 7.5, 30°C | Xanthobacter autotrophicus | |
28 | - |
(S)-2-butanol | wild-type, pH 7.5, 30°C | Xanthobacter autotrophicus | |
68 | - |
(R)-2-butanol | wild-type, pH 7.5, 30°C | Xanthobacter autotrophicus | |
72 | - |
2-butanone | mutant K214A, pH 7.5, 30°C | Xanthobacter autotrophicus | |
120 | - |
2-butanone | wild-type, pH 7.5, 30°C | Xanthobacter autotrophicus | |
180 | - |
2-butanone | mutant R211A, pH 7.5, 30°C | Xanthobacter autotrophicus | |
720 | - |
2-propanol | mutant R211A, pH 7.5, 30°C | Xanthobacter autotrophicus | |
950 | - |
2-propanol | mutant K214A, pH 7.5, 30°C | Xanthobacter autotrophicus | |
1400 | - |
2-propanol | wild-type, pH 7.5, 30°C | Xanthobacter autotrophicus |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Xanthobacter autotrophicus | - |
- |
- |
Xanthobacter autotrophicus | Q56841 | - |
- |
Purification (Comment) | Organism |
---|---|
recombinant enzyme | Xanthobacter autotrophicus |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
(R)-2-butanol + NAD+ | - |
Xanthobacter autotrophicus | 2-butanone + NADH + H+ | - |
r | |
(S)-2-butanol + NAD+ | - |
Xanthobacter autotrophicus | 2-butanone + NADH + H+ | - |
r | |
2-(2-hydroxyethylthio)ethanesulfonate + NAD+ | achiral mimic of both R-hydroxypropyl-CoM and S-hydroxypropyl-CoM, substrate for both the R- and S-HPCDH enzymes with identical Km values | Xanthobacter autotrophicus | 2-(formylmethylthio)ethanesulfonate + NADH + H+ | - |
r | |
2-(R)-hydroxypropyl-CoM + NAD+ | - |
Xanthobacter autotrophicus | 2-oxopropyl-CoM + NADH + H+ | - |
r | |
2-(R)-hydroxypropyl-CoM + NAD+ | poor substrate. S-HPCDH3 cannot bind hydroxypropyl-CoM with CoM oriented properly in the sulfonate-binding pocket that consists of residues R211 and K214. R-hydroxypropyl-CoM binds to S-HPCDH3 with a 290-fold lower affinity but in an orientation where the hydroxyl and hydrogen on C2 can be more properly aligned with tyrosine 156 and NAD+, such that kcat decreases by 4.5-fold relative to the natural substrate S-hydroxypropyl-CoM | Xanthobacter autotrophicus | 2-oxopropyl-CoM + NADH + H+ | - |
r | |
2-(S)-hydroxypropyl-CoM + NAD+ | - |
Xanthobacter autotrophicus | 2-oxopropyl-CoM + NADH + H+ | - |
r | |
2-(S)-hydroxypropyl-CoM + NAD+ | oxidation of S-hydroxypropyl-CoM with a kcat that is 402 times less than that for R-hydroxypropyl-CoM | Xanthobacter autotrophicus | 2-oxopropyl-CoM + NADH + H+ | - |
r | |
2-butanone + NADH + H+ | - |
Xanthobacter autotrophicus | (S)-2-butanol + (R)-2-butanol + NAD+ | - |
r | |
2-butanone + NADH + H+ | - |
Xanthobacter autotrophicus | (S)-2-butanol + (R)-2-butanol + NAD+ | without additions, 99.2% (S)-enantiomer + 0.8% (R)-enantiomer, in presence of 1 mM ethansulfonate 99.0% (S)-enantiomer + 2% (R)-enantiomer. Mutant K214A, without additions, 91.6% (S)-enantiomer + 8.4% (R)-enantiomer, in presence of 1 mM ethansulfonate 90.9% (S)-enantiomer + 9.1% (R)-enantiomer | r | |
2-oxopropyl-CoM + NADH + H+ | - |
Xanthobacter autotrophicus | 2-(R)-hydroxypropyl-CoM + NAD+ | - |
r | |
2-propanol + NAD+ | - |
Xanthobacter autotrophicus | acetone + NADH + H+ | - |
r |
Synonyms | Comment | Organism |
---|---|---|
HPCDH3 | - |
Xanthobacter autotrophicus |
xecE1 | - |
Xanthobacter autotrophicus |
xecE3 | - |
Xanthobacter autotrophicus |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.013 | - |
2-(S)-hydroxypropyl-CoM | mutant S143A, pH 7.5, 30°C | Xanthobacter autotrophicus | |
0.039 | - |
2-butanone | mutant K214A, pH 7.5, 30°C | Xanthobacter autotrophicus | |
0.044 | - |
2-butanone | wild-type, pH 7.5, 30°C | Xanthobacter autotrophicus | |
0.072 | - |
2-butanone | mutant R211A, pH 7.5, 30°C | Xanthobacter autotrophicus | |
0.12 | - |
2-(S)-hydroxypropyl-CoM | pH 7.5, 30°C | Xanthobacter autotrophicus | |
0.65 | - |
2-(S)-hydroxypropyl-CoM | mutant Y156A, pH 7.5, 30°C | Xanthobacter autotrophicus | |
1 | - |
(R)-2-butanol | wild-type, pH 7.5, 30°C | Xanthobacter autotrophicus | |
1.7 | - |
2-propanol | mutant K214A, pH 7.5, 30°C | Xanthobacter autotrophicus | |
1.7 | - |
2-propanol | mutant R211A, pH 7.5, 30°C | Xanthobacter autotrophicus | |
2 | - |
2-propanol | wild-type, pH 7.5, 30°C | Xanthobacter autotrophicus | |
2.6 | - |
(S)-2-butanol | wild-type, pH 7.5, 30°C | Xanthobacter autotrophicus | |
3.8 | - |
2-(2-hydroxyethylthio)ethanesulfonate | wild-type, pH 7.5, 30°C | Xanthobacter autotrophicus | |
5.5 | - |
2-(R)-hydroxypropyl-CoM | wild-type, pH 7.5, 30°C | Xanthobacter autotrophicus | |
5.8 | - |
2-(S)-hydroxypropyl-CoM | mutant K214A, pH 7.5, 30°C | Xanthobacter autotrophicus | |
8.6 | - |
2-oxopropyl-CoM | mutant R211A, pH 7.5, 30°C | Xanthobacter autotrophicus | |
9.6 | - |
2-oxopropyl-CoM | mutant K214A, pH 7.5, 30°C | Xanthobacter autotrophicus | |
11 | - |
2-oxopropyl-CoM | wild-type, pH 7.5, 30°C | Xanthobacter autotrophicus | |
16 | - |
2-(S)-hydroxypropyl-CoM | mutant R211A, pH 7.5, 30°C | Xanthobacter autotrophicus | |
25 | - |
2-(S)-hydroxypropyl-CoM | wild-type, pH 7.5, 30°C | Xanthobacter autotrophicus | |
29 | - |
2-oxopropyl-CoM | pH 7.5, 30°C | Xanthobacter autotrophicus | |
49 | - |
2-(R)-hydroxypropyl-CoM | pH 7.5, 30°C | Xanthobacter autotrophicus |
Ki Value [mM] | Ki Value maximum [mM] | Inhibitor | Comment | Organism | Structure |
---|---|---|---|---|---|
0.29 | - |
2-(2-methyl-2-hydroxypropylthio)ethanesulfonate | pH 7.5, 30°C | Xanthobacter autotrophicus | |
0.29 | - |
2-(2-methyl-2-hydroxypropylthio)ethanesulfonate | wild-type, pH 7.5, 30°C | Xanthobacter autotrophicus |
kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.0014 | - |
2-(S)-hydroxypropyl-CoM | mutant S143A, pH 7.5, 30°C | Xanthobacter autotrophicus | |
0.0148 | - |
(R)-2-butanol | wild-type, pH 7.5, 30°C | Xanthobacter autotrophicus | |
0.0928 | - |
(S)-2-butanol | wild-type, pH 7.5, 30°C | Xanthobacter autotrophicus | |
0.35 | - |
2-(S)-hydroxypropyl-CoM | mutant Y156A, pH 7.5, 30°C | Xanthobacter autotrophicus | |
0.37 | - |
2-butanone | wild-type, pH 7.5, 30°C | Xanthobacter autotrophicus | |
0.4 | - |
2-butanone | mutant R211A, pH 7.5, 30°C | Xanthobacter autotrophicus | |
0.53 | - |
2-(S)-hydroxypropyl-CoM | pH 7.5, 30°C | Xanthobacter autotrophicus | |
0.54 | - |
2-butanone | mutant K214A, pH 7.5, 30°C | Xanthobacter autotrophicus | |
0.6 | - |
2-(R)-hydroxypropyl-CoM | wild-type, pH 7.5, 30°C | Xanthobacter autotrophicus | |
0.75 | - |
2-oxopropyl-CoM | mutant R211A, pH 7.5, 30°C | Xanthobacter autotrophicus | |
0.81 | - |
2-oxopropyl-CoM | mutant K214A, pH 7.5, 30°C | Xanthobacter autotrophicus | |
1.4 | - |
2-propanol | wild-type, pH 7.5, 30°C | Xanthobacter autotrophicus | |
1.8 | - |
2-propanol | mutant K214A, pH 7.5, 30°C | Xanthobacter autotrophicus | |
2.4 | - |
2-propanol | mutant R211A, pH 7.5, 30°C | Xanthobacter autotrophicus | |
2.5 | - |
2-(S)-hydroxypropyl-CoM | mutant K214A, pH 7.5, 30°C | Xanthobacter autotrophicus | |
10 | - |
2-(S)-hydroxypropyl-CoM | mutant R211A, pH 7.5, 30°C | Xanthobacter autotrophicus | |
39 | - |
2-oxopropyl-CoM | wild-type, pH 7.5, 30°C | Xanthobacter autotrophicus | |
420 | - |
2-oxopropyl-CoM | pH 7.5, 30°C | Xanthobacter autotrophicus | |
500 | - |
2-(R)-hydroxypropyl-CoM | pH 7.5, 30°C | Xanthobacter autotrophicus | |
790 | - |
2-(S)-hydroxypropyl-CoM | wild-type, pH 7.5, 30°C | Xanthobacter autotrophicus |