Activating Compound | Comment | Organism | Structure |
---|---|---|---|
ethanesulfonate | the addition of ethanesulfonate dramatically increases the enentiomeric excess of (S)-2-butanol produced by all forms of recombinant R-HPCDH except the R152A and R196A mutants. The effect of ethanesulfonate and other additives on (S)-butanol production is termed enantioselective modulation, and it is highly selective for sulfonate containing molecules. The interpretation is that the sulfonate of ethanesulfonate interacts with R152 and R196 in the CoM binding pocket alongside 2-butanone, in a way that discourages a si-face hydride addition to produce (R)-2-butanol | Xanthobacter autotrophicus |
Crystallization (Comment) | Organism |
---|---|
purified recombinant enzyme, X-ray diffraction structure determination and analysis | Xanthobacter autotrophicus |
Protein Variants | Comment | Organism |
---|---|---|
additional information | comparison of steady-state kinetics and 2-butanol production arte and stereochemistry of wild-type and mutant enzymes, overview | Xanthobacter autotrophicus |
R152A | site-directed mutagenesis, the mutant shows altered substrate specificity and interaction with additives compared to wild-type enzyme | Xanthobacter autotrophicus |
R196A | site-directed mutagenesis, the mutant shows altered substrate specificity and interaction with additives compared to wild-type enzyme | Xanthobacter autotrophicus |
R203A | site-directed mutagenesis, the mutant shows altered substrate specificity and interaction with additives compared to wild-type enzyme | Xanthobacter autotrophicus |
R209A | site-directed mutagenesis, the mutant shows altered substrate specificity and interaction with additives compared to wild-type enzyme | Xanthobacter autotrophicus |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
(2S)-2-hydroxypropyl-CoM | competitive inhibition | Xanthobacter autotrophicus | |
2-(2-methyl-2-hydroxypropylthio)-ethanesulfonate | M-HPC, competitive inhibition | Xanthobacter autotrophicus | |
CoM | mixed inhibition | Xanthobacter autotrophicus |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
additional information | - |
additional information | steady-state kinetics, comparison of steady-state kinetics with wild-type an d mutant enzymes, overview | Xanthobacter autotrophicus | |
0.1 | - |
(2S)-2-hydroxypropyl-CoM | recombinant enzyme, pH and temperature not specified in the publication | Xanthobacter autotrophicus | |
0.102 | - |
(2R)-2-hydroxypropyl-CoM | recombinant enzyme, pH and temperature not specified in the publication | Xanthobacter autotrophicus | |
0.75 | - |
2-(2-hydroxyethylthio)ethanesulfonate | recombinant enzyme, pH and temperature not specified in the publication | Xanthobacter autotrophicus | |
1.08 | - |
(2R)-2-octanol | recombinant enzyme, pH and temperature not specified in the publication | Xanthobacter autotrophicus | |
2.64 | - |
(2R)-2-heptanol | recombinant enzyme, pH and temperature not specified in the publication | Xanthobacter autotrophicus | |
4.6 | - |
(2R)-2-hexanol | recombinant enzyme, pH and temperature not specified in the publication | Xanthobacter autotrophicus | |
20 | - |
(2R)-2-pentanol | recombinant enzyme, pH and temperature not specified in the publication | Xanthobacter autotrophicus | |
153 | - |
(2S)-2-pentanol | recombinant enzyme, pH and temperature not specified in the publication | Xanthobacter autotrophicus | |
315 | - |
(2S)-2-butanol | recombinant enzyme, pH and temperature not specified in the publication | Xanthobacter autotrophicus | |
326 | - |
(2R)-2-butanol | recombinant enzyme, pH and temperature not specified in the publication | Xanthobacter autotrophicus | |
1726 | - |
2-propanol | recombinant enzyme, pH and temperature not specified in the publication | Xanthobacter autotrophicus |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
(2R)-2-hydroxypropyl-CoM + NAD+ | Xanthobacter autotrophicus | - |
2-oxopropyl-CoM + NADH + H+ | - |
r | |
(2R)-2-hydroxypropyl-CoM + NAD+ | Xanthobacter autotrophicus ATCC BAA-1158 | - |
2-oxopropyl-CoM + NADH + H+ | - |
r |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Xanthobacter autotrophicus | Q56840 | - |
- |
Xanthobacter autotrophicus ATCC BAA-1158 | Q56840 | - |
- |
Reaction | Comment | Organism | Reaction ID |
---|---|---|---|
2-(R)-hydroxypropyl-CoM + NAD+ = 2-oxopropyl-CoM + NADH + H+ | active site structure modeling and stereochemistry of reaction mechanism, overview | Xanthobacter autotrophicus |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
(2R)-2-butanol + NAD+ | - |
Xanthobacter autotrophicus | 2-butanone + NADH + H+ | - |
r | |
(2R)-2-heptanol + NAD+ | - |
Xanthobacter autotrophicus | 2-heptanone + NADH + H+ | - |
r | |
(2R)-2-hexanol + NAD+ | - |
Xanthobacter autotrophicus | 2-hexanone + NADH + H+ | - |
r | |
(2R)-2-hydroxypropyl-CoM + NAD+ | - |
Xanthobacter autotrophicus | 2-oxopropyl-CoM + NADH + H+ | - |
r | |
(2R)-2-hydroxypropyl-CoM + NAD+ | substrate binding structure, overview | Xanthobacter autotrophicus | 2-oxopropyl-CoM + NADH + H+ | - |
r | |
(2R)-2-hydroxypropyl-CoM + NAD+ | - |
Xanthobacter autotrophicus ATCC BAA-1158 | 2-oxopropyl-CoM + NADH + H+ | - |
r | |
(2R)-2-hydroxypropyl-CoM + NAD+ | substrate binding structure, overview | Xanthobacter autotrophicus ATCC BAA-1158 | 2-oxopropyl-CoM + NADH + H+ | - |
r | |
(2R)-2-octanol + NAD+ | - |
Xanthobacter autotrophicus | 2-octanone + NADH + H+ | - |
r | |
(2R)-2-pentanol + NAD+ | - |
Xanthobacter autotrophicus | 2-pentanone + NADH + H+ | - |
r | |
(2S)-2-butanol + NAD+ | very low activity | Xanthobacter autotrophicus | 2-butanone + NADH + H+ | - |
r | |
(2S)-2-hydroxypropyl-CoM + NAD+ | competitive inhibitor to the R-enantiomer of substrate, very low activity | Xanthobacter autotrophicus | 2-oxopropyl-CoM + NADH + H+ | - |
r | |
(2S)-2-hydroxypropyl-CoM + NAD+ | competitive inhibitor to the R-enantiomer of substrate, very low activity | Xanthobacter autotrophicus ATCC BAA-1158 | 2-oxopropyl-CoM + NADH + H+ | - |
r | |
(2S)-2-pentanol + NAD+ | - |
Xanthobacter autotrophicus | 2-pentanone + NADH + H+ | - |
r | |
2-(2-hydroxyethylthio)ethanesulfonate + NADH + H+ | very low activity | Xanthobacter autotrophicus | ? | - |
? | |
2-butanone + NADH + H+ | with no additives present, all forms of recombinant R-HPCDH prefer a re-face hydride addition to produce an enantiomeric excess (EE) of (S)-2-butanol, S- and R-2-butanol are comparably good substrates for the reverse reaction. The sulfonate of ethanesulfonate interacts with R152 and R196 in the CoM binding pocket alongside 2-butanone, in a way that discourages a si-face hydride addition to produce (R)-2-butanol | Xanthobacter autotrophicus | (2R)-2-butanol + NAD+ | - |
r | |
2-butanone + NADH + H+ | with no additives present, all forms of recombinant R-HPCDH prefer a re-face hydride addition to produce an enantiomeric excess (EE) of (S)-2-butanol, S- and R-2-butanol are comparably good substrates for the reverse reaction. The sulfonate of ethanesulfonate interacts with R152 and R196 in the CoM binding pocket alongside 2-butanone, in a way that discourages a si-face hydride addition to produce (R)-2-butanol | Xanthobacter autotrophicus | (2S)-2-butanol + NAD+ | - |
r | |
2-propanol + NAD+ | - |
Xanthobacter autotrophicus | 2-propanone + NADH + H+ | - |
r | |
2-propanol + NAD+ | - |
Xanthobacter autotrophicus ATCC BAA-1158 | 2-propanone + NADH + H+ | - |
r | |
additional information | substrate specificity of the stereochemically different isozymes, R- and S-HPCDH (EC 1.1.1.269) are 41% identical to each other, overview. No activity with (S)-2-hexanol, (S)-2-heptanol, and (S)-2-octanol. Stereochemistry of products from reaction with wild-type and mutant enzymes with or without addition of additives, i.e. CH3CH2SO3- Na+, HSCH2CH2SO3- Na+, CH3CH2COO- Na+, CH3COO- Na+, CH3CH2NH3 +Cl-, CH3CH2OH, Na2SO4, and NaCl, overview | Xanthobacter autotrophicus | ? | - |
- |
|
additional information | substrate specificity of the stereochemically different isozymes, R- and S-HPCDH (EC 1.1.1.269) are 41% identical to each other, overview. No activity with (S)-2-hexanol, (S)-2-heptanol, and (S)-2-octanol. Stereochemistry of products from reaction with wild-type and mutant enzymes with or without addition of additives, i.e. CH3CH2SO3- Na+, HSCH2CH2SO3- Na+, CH3CH2COO- Na+, CH3COO- Na+, CH3CH2NH3 +Cl-, CH3CH2OH, Na2SO4, and NaCl, overview | Xanthobacter autotrophicus ATCC BAA-1158 | ? | - |
- |
Synonyms | Comment | Organism |
---|---|---|
(R)-hydroxypropyl-coenzyme M dehydrogenase | - |
Xanthobacter autotrophicus |
R-HPCDH | - |
Xanthobacter autotrophicus |
xecD | - |
Xanthobacter autotrophicus |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.044 | - |
(2S)-2-hydroxypropyl-CoM | recombinant enzyme, pH and temperature not specified in the publication | Xanthobacter autotrophicus | |
0.29 | - |
2-(2-hydroxyethylthio)ethanesulfonate | recombinant enzyme, pH and temperature not specified in the publication | Xanthobacter autotrophicus | |
0.98 | - |
(2R)-2-butanol | recombinant enzyme, pH and temperature not specified in the publication | Xanthobacter autotrophicus | |
1.1 | - |
(2R)-2-pentanol | recombinant enzyme, pH and temperature not specified in the publication | Xanthobacter autotrophicus | |
1.7 | - |
(2R)-2-octanol | recombinant enzyme, pH and temperature not specified in the publication | Xanthobacter autotrophicus | |
1.8 | - |
(2R)-2-heptanol | recombinant enzyme, pH and temperature not specified in the publication | Xanthobacter autotrophicus | |
2.2 | - |
(2S)-2-butanol | recombinant enzyme, pH and temperature not specified in the publication | Xanthobacter autotrophicus | |
2.7 | - |
(2R)-2-hexanol | recombinant enzyme, pH and temperature not specified in the publication | Xanthobacter autotrophicus | |
2.9 | - |
2-propanol | recombinant enzyme, pH and temperature not specified in the publication | Xanthobacter autotrophicus | |
2.9 | - |
(2S)-2-pentanol | recombinant enzyme, pH and temperature not specified in the publication | Xanthobacter autotrophicus | |
26.8 | - |
(2R)-2-hydroxypropyl-CoM | recombinant enzyme, pH and temperature not specified in the publication | Xanthobacter autotrophicus |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
NAD+ | - |
Xanthobacter autotrophicus | |
NADH | - |
Xanthobacter autotrophicus |
Ki Value [mM] | Ki Value maximum [mM] | Inhibitor | Comment | Organism | Structure |
---|---|---|---|---|---|
0.156 | - |
(2S)-2-hydroxypropyl-CoM | pH and temperature not specified in the publication, versus (2R)-2-hydroxypropyl-CoM | Xanthobacter autotrophicus | |
0.406 | - |
2-(2-methyl-2-hydroxypropylthio)-ethanesulfonate | pH and temperature not specified in the publication, versus (2R)-2-hydroxypropyl-CoM | Xanthobacter autotrophicus | |
2.25 | - |
CoM | pH and temperature not specified in the publication, competitive versus (2R)-2-hydroxypropyl-CoM | Xanthobacter autotrophicus | |
3.62 | - |
CoM | pH and temperature not specified in the publication, uncompetitive | Xanthobacter autotrophicus |
General Information | Comment | Organism |
---|---|---|
evolution | the enzyme belongs to the short-chain dehydrogenases/reductase (SDR) superfamily of enzymes. The C-terminal domains of SDR enzymes are responsible for imparting substrate specificity | Xanthobacter autotrophicus |
malfunction | substitution of R152 or R196 for alanine inhibits ethanesulfonate binding to the extent that its addition does not increase the EE of (S)-2-butanol produced by these mutants | Xanthobacter autotrophicus |
metabolism | (R)- and (S)-hydroxypropyl-coenzyme M dehydrogenase (R- and S-HPCDH), are part of a bacterial pathway of short-chain alkene and epoxide metabolism. R- and S-HPCDH act on different substrate enantiomers in a common pathway | Xanthobacter autotrophicus |
additional information | structure-function relationship, active site structure modeling and stereochemistry of reaction mechanism, overview. The C-terminal domains of SDR enzymes are responsible for imparting substrate specificity. Two arginine residues, R152 and R196, play a key role in substrate binding and stereoselectivity of enzyme R-HPCDH. R152 and R196 bind the sulfonate of 2-oxopropyl-CoM (2-KPC) | Xanthobacter autotrophicus |
kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.0017 | - |
2-propanol | recombinant enzyme, pH and temperature not specified in the publication | Xanthobacter autotrophicus | |
0.003 | - |
(2R)-2-butanol | recombinant enzyme, pH and temperature not specified in the publication | Xanthobacter autotrophicus | |
0.007 | - |
(2S)-2-butanol | recombinant enzyme, pH and temperature not specified in the publication | Xanthobacter autotrophicus | |
0.019 | - |
(2S)-2-pentanol | recombinant enzyme, pH and temperature not specified in the publication | Xanthobacter autotrophicus | |
0.055 | - |
(2R)-2-pentanol | recombinant enzyme, pH and temperature not specified in the publication | Xanthobacter autotrophicus | |
0.39 | - |
2-(2-hydroxyethylthio)ethanesulfonate | recombinant enzyme, pH and temperature not specified in the publication | Xanthobacter autotrophicus | |
0.44 | - |
(2S)-2-hydroxypropyl-CoM | recombinant enzyme, pH and temperature not specified in the publication | Xanthobacter autotrophicus | |
0.59 | - |
(2R)-2-hexanol | recombinant enzyme, pH and temperature not specified in the publication | Xanthobacter autotrophicus | |
0.68 | - |
(2R)-2-heptanol | recombinant enzyme, pH and temperature not specified in the publication | Xanthobacter autotrophicus | |
1.57 | - |
(2R)-2-octanol | recombinant enzyme, pH and temperature not specified in the publication | Xanthobacter autotrophicus | |
262.75 | - |
(2R)-2-hydroxypropyl-CoM | recombinant enzyme, pH and temperature not specified in the publication | Xanthobacter autotrophicus |