EC Number | Activating Compound | Comment | Organism | Structure |
---|---|---|---|---|
1.1.1.2 | Isopropanol | highly efficient reduction of ketones by coupling with excess isopropanol to regenerate NADPH | Thermococcus guaymasensis |
EC Number | Cloned (Comment) | Organism |
---|---|---|
1.1.1.2 | the gene encoding the enzyme is cloned and sequenced, it shows high sequence homology to zinc-containing alcohol dehydrogenases and L-threonine dehydrogenases with binding motifs of catalytic zinc and NADP+ | Thermococcus guaymasensis |
EC Number | Inhibitors | Comment | Organism | Structure |
---|---|---|---|---|
1.1.1.2 | Cd2+ | 1 mM, complete inhibition | Thermococcus guaymasensis | |
1.1.1.2 | Cu2+ | 1 mM, complete inhibition | Thermococcus guaymasensis | |
1.1.1.2 | EDTA | 1 mM, slightly decreased the enzyme activity | Thermococcus guaymasensis | |
1.1.1.2 | Fe2+ | 1 mM, 10-20% decrease in activity | Thermococcus guaymasensis | |
1.1.1.2 | Hg2+ | 1 mM, complete inhibition | Thermococcus guaymasensis | |
1.1.1.2 | Ni2+ | 1 mM, 10-20% decrease in activity | Thermococcus guaymasensis | |
1.1.1.2 | Zn2+ | 1 mM, complete inhibition | Thermococcus guaymasensis |
EC Number | KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|---|
1.1.1.2 | 0.011 | - |
NADPH | cosubstrate: 2-butanone, pH 7.5, 80°C | Thermococcus guaymasensis | |
1.1.1.2 | 0.21 | - |
diacetyl | cosubstrate: NADPH, pH 7.5, 80°C | Thermococcus guaymasensis | |
1.1.1.2 | 0.31 | - |
2-butanone | cosubstrate: NADPH, pH 7.5, 80°C | Thermococcus guaymasensis | |
1.1.1.2 | 0.32 | - |
acetoin | cosubstrate: NADPH, pH 7.5, 80°C | Thermococcus guaymasensis | |
1.1.1.2 | 0.38 | - |
2-butanol | cosubstrate: NADP+, pH 7.5, 80°C | Thermococcus guaymasensis | |
1.1.1.2 | 0.4 | - |
NADP+ | cosubstrate: 2-butanol, pH 7.5, 80°C | Thermococcus guaymasensis | |
1.1.1.2 | 15.2 | - |
(2R,3R)-2,3-butanediol | cosubstrate: NADP+, pH 7.5, 80°C | Thermococcus guaymasensis | |
1.1.1.2 | 19.3 | - |
meso-2,3-butanediol | cosubstrate: NADP+, pH 7.5, 80°C | Thermococcus guaymasensis | |
1.1.1.2 | 246 | - |
(2S,3S)-2,3-butanediol | cosubstrate: NADP+, pH 7.5, 80°C | Thermococcus guaymasensis |
EC Number | Metals/Ions | Comment | Organism | Structure |
---|---|---|---|---|
1.1.1.2 | Ca2+ | 1 mM, less than 15% increase of activity | Thermococcus guaymasensis | |
1.1.1.2 | Co2+ | 1 mM, less than 15% increase of activity | Thermococcus guaymasensis | |
1.1.1.2 | Mg2+ | 1 mM, less than 15% increase of activity | Thermococcus guaymasensis | |
1.1.1.2 | Mn2+ | 1 mM, less than 15% increase of activity | Thermococcus guaymasensis | |
1.1.1.2 | Zinc | contains 0.9 gatoms of zinc per subunit | Thermococcus guaymasensis |
EC Number | Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
---|---|---|---|---|
1.1.1.2 | 40000 | - |
4 * 40000, SDS-PAGE | Thermococcus guaymasensis |
1.1.1.2 | 135000 | - |
gel filtration | Thermococcus guaymasensis |
EC Number | Organic Solvent | Comment | Organism |
---|---|---|---|
1.1.1.2 | isopropanol | solvent-tolerant enzyme, highly efficient reduction of ketones by coupling with excess isopropanol to regenerate NADPH | Thermococcus guaymasensis |
1.1.1.2 | Methanol | the purified enzyme does not oxidize methanol. The enzyme retains almost its full activity when the concentration of methanol is up to 5% (v/v). When the concentration of methanol is 40% (v/v) in the assay mixture, the enzyme activities remain at 30% and 15% of the full activity on both oxidation and reduction, respectively | Thermococcus guaymasensis |
EC Number | Organism | UniProt | Comment | Textmining |
---|---|---|---|---|
1.1.1.2 | Thermococcus guaymasensis | F8S123 | - |
- |
EC Number | Oxidation Stability | Organism |
---|---|---|
1.1.1.2 | the enzyme is oxygen sensitive, although it is much more resistant to oxidation than iron-containing alcohol dehydrogenases. The time required to decrease 50% of the full activity upon exposure to the air (t1/2) is about 4 h, and such inactivation is slightly decreased in the presence of 2 mM dithiothreitol | Thermococcus guaymasensis |
EC Number | Purification (Comment) | Organism |
---|---|---|
1.1.1.2 | - |
Thermococcus guaymasensis |
EC Number | Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|---|
1.1.1.2 | (2R,3R)-2,3-butanediol + NADP+ | the enzyme shows higher oxidation activities on the (2R,3R)-(-)-2,3-butanediol and meso-2,3-butanediol than on the (2S,3S)-(+)-2,3-butanediol, indicating that the enzyme predominantly oxidizes the R-hydroxyl group of 2,3-butanediol and minorly functions on the S-hydroxyl group. When meso-2,3-butanediol is oxidized, (S)-2-acetoin is the predominant product, with an enantiomeric excess (ee) of 88%, while oxidation of (2R,3R)-2,3-butanediol results in the production of (R)-2-acetoin with higher stereoselectivity (94% ee) | Thermococcus guaymasensis | (R)-2-acetoin + NADPH + H+ | - |
r | |
1.1.1.2 | (2S,3S)-2,3-butanediol + NADP+ | the enzyme shows higher oxidation activities on the (2R,3R)-(-)-2,3-butanediol and meso-2,3-butanediol than on the (2S,3S)-(+)-2,3-butanediol, indicating that the enzyme predominantly oxidizes the R-hydroxyl group of 2,3-butanediol and minorly functions on the S-hydroxyl group. When meso-2,3-butanediol is oxidized, (S)-2-acetoin is the predominant product, with an enantiomeric excess (ee) of 88%, while oxidation of meso-2,3-butanediol results in the production of (R)-2-acetoin with higher stereoselectivity (94% ee) | Thermococcus guaymasensis | (S)-2-acetoin + NADPH + H+ | - |
r | |
1.1.1.2 | 1,2-butanediol + NADP+ | 25.6% of the activity compared to the substrate 2-butanol | Thermococcus guaymasensis | ? + NADPH + H+ | - |
? | |
1.1.1.2 | 1,3-butanediol + NADP+ | 35.4% of the activity compared to the substrate 2-butanol | Thermococcus guaymasensis | ? + NADPH + H+ | - |
? | |
1.1.1.2 | 1-butanol + NADP+ | 5.1% of the activity compared to the substrate 2-butanol | Thermococcus guaymasensis | butyraldehyde + NADPH + H+ | - |
r | |
1.1.1.2 | 1-propanol + NADP+ | 15.1% of the activity compared to the substrate 2-butanol | Thermococcus guaymasensis | propanal + NADPH + H+ | - |
? | |
1.1.1.2 | 2,4-pentanediol + NADP+ | 9.2% of the activity compared to the substrate 2-butanol | Thermococcus guaymasensis | ? + NADPH + H+ | - |
? | |
1.1.1.2 | 2-butanol + NADP+ | the enzyme does not possess stereoselectivity on 2-butanol or 2-butanone as substrate | Thermococcus guaymasensis | 2-butanone + NADPH + H+ | - |
r | |
1.1.1.2 | 2-butanone + NADPH + H+ | the enzyme does not possess stereoselectivity on 2-butanol or 2-butanone as substrate | Thermococcus guaymasensis | 2-butanol + NADP+ | - |
r | |
1.1.1.2 | 2-pentanol + NADP+ | 66.3% of the activity compared to the substrate 2-butanol | Thermococcus guaymasensis | 2-pentanone + NADPH + H+ | - |
r | |
1.1.1.2 | 2-pentanone + NADPH + H+ | 86.2% of the activity compared to the substrate 2-butanone | Thermococcus guaymasensis | 2-pentanol + NADP+ | - |
r | |
1.1.1.2 | 2-propanol + NADP+ | 88% of the activity compared to the substrate 2-butanol | Thermococcus guaymasensis | acetone + NADPH + H+ | - |
r | |
1.1.1.2 | acetaldehyde + NADPH + H+ | - |
Thermococcus guaymasensis | ethanol + NADP+ | - |
r | |
1.1.1.2 | acetoin + NADPH + H+ | 93% of the activity compared to the substrate 2-butanone. The racemic R/S-acetoin is used since neither R- nor S-acetoin is commercially available. Consistently, the reduction of racemic R/S-acetoin formed (2R,3R)-2,3-butanediol (presumably from R-acetoin) and meso-2,3-butanediol (presumably from S-acetoin) with extremely high stereoselectivity (99% ee over (2S,3S)-(+)-2,3-butanediol) | Thermococcus guaymasensis | 2,3-butanediol + NADP+ | - |
r | |
1.1.1.2 | acetone + NADPH + H+ | 149% of the activity compared to the substrate 2-butanone | Thermococcus guaymasensis | 2-propanol + NADP+ | - |
r | |
1.1.1.2 | butyraldehyde + NADPH + H+ | 112.4% of the activity compared to the substrate 2-butanone | Thermococcus guaymasensis | 1-butanol + NADP+ | - |
r | |
1.1.1.2 | diacetyl + NADPH + H+ | the enzyme can not oxidize acetoin to diacetyl | Thermococcus guaymasensis | acetoin + NADP+ | - |
ir | |
1.1.1.2 | ethanol + NADP+ | 5.7% of the activity compared to the substrate 2-butanol | Thermococcus guaymasensis | acetaldehyde + NADPH + H+ | - |
r | |
1.1.1.2 | meso-2,3-butanediol + NADP+ | the enzyme shows higher oxidation activities on the (2R,3R)-(-)-2,3-butanediol and meso-2,3-butanediol than on the (2S,3S)-(+)-2,3-butanediol, indicating that the enzyme predominantly oxidizes the R-hydroxyl group of 2,3-butanediol and minorly functions on the S-hydroxyl group. When meso-2,3-butanediol is oxidized, (S)-2-acetoin is the predominant product, with an enantiomeric excess (ee) of 88%, while oxidation of meso-2,3-butanediol results in the production of (R)-2-acetoin with higher stereoselectivity (94% ee) | Thermococcus guaymasensis | (S)-2-acetoin + NADPH + H+ | - |
r | |
1.1.1.2 | additional information | the enzyme shows higher activities with the use of secondary alcohols such as 2-butanol and 2-pentanol as substrates, suggesting that the enzyme is a primary-secondary alcohol dehydrogenase. Alcohols oxidized at an acticity of less than 5% compared to the substrate 2-butanol: 1-pentanol, glycerol, 1,2-pentanediol. No oxidation of methanol, 1,4-butanediol, 1,5-pentanediol, ethanediol, phenylethanol, L-serine, L-threonine, acetoin | Thermococcus guaymasensis | ? | - |
? |
EC Number | Subunits | Comment | Organism |
---|---|---|---|
1.1.1.2 | homotetramer | 4 * 40000, SDS-PAGE | Thermococcus guaymasensis |
EC Number | Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|---|
1.1.1.2 | 95 | - |
activity increases along with the elevated temperatures up to 95°C | Thermococcus guaymasensis |
EC Number | Temperature Stability Minimum [°C] | Temperature Stability Maximum [°C] | Comment | Organism |
---|---|---|---|---|
1.1.1.2 | 80 | - |
t1/2: 70 h | Thermococcus guaymasensis |
1.1.1.2 | 95 | - |
t1/2: 24 h | Thermococcus guaymasensis |
EC Number | Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|---|
1.1.1.2 | 142 | - |
acetoin | cosubstrate: NADPH, pH 7.5, 80°C | Thermococcus guaymasensis | |
1.1.1.2 | 158 | - |
NADPH | cosubstrate: 2-butanone, pH 7.5, 80°C | Thermococcus guaymasensis | |
1.1.1.2 | 190 | - |
2-butanone | cosubstrate: NADPH, pH 7.5, 80°C | Thermococcus guaymasensis | |
1.1.1.2 | 202 | - |
diacetyl | cosubstrate: NADPH, pH 7.5, 80°C | Thermococcus guaymasensis | |
1.1.1.2 | 512 | - |
(2S,3S)-2,3-butanediol | cosubstrate: NADP+, pH 7.5, 80°C | Thermococcus guaymasensis | |
1.1.1.2 | 740 | - |
(2R,3R)-2,3-butanediol | cosubstrate: NADP+, pH 7.5, 80°C | Thermococcus guaymasensis | |
1.1.1.2 | 833 | - |
2-butanol | cosubstrate: NADP+, pH 7.5, 80°C | Thermococcus guaymasensis | |
1.1.1.2 | 952 | - |
meso-2,3-butanediol | cosubstrate: NADP+, pH 7.5, 80°C | Thermococcus guaymasensis | |
1.1.1.2 | 1333 | - |
NADP+ | cosubstrate: 2-butanol, pH 7.5, 80°C | Thermococcus guaymasensis |
EC Number | pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|---|
1.1.1.2 | 7.5 | - |
2-butanone reduction | Thermococcus guaymasensis |
1.1.1.2 | 10.5 | - |
2-butanol oxidation | Thermococcus guaymasensis |
EC Number | Cofactor | Comment | Organism | Structure |
---|---|---|---|---|
1.1.1.2 | NADP+ | Km values and catalytic efficiency for NADPH are 40times lower and 5times higher than those for NADP+, respectively | Thermococcus guaymasensis | |
1.1.1.2 | NADPH | Km values and catalytic efficiency for NADPH are 40times lower and 5times higher than those for NADP+, respectively | Thermococcus guaymasensis |
EC Number | General Information | Comment | Organism |
---|---|---|---|
1.1.1.2 | physiological function | the physiological roles of the enzyme are proposed to be in the formation of alcohols such as ethanol or acetoin concomitant to the NADPH oxidation | Thermococcus guaymasensis |
EC Number | kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|---|
1.1.1.2 | 2.08 | - |
(2S,3S)-2,3-butanediol | cosubstrate: NADP+, pH 7.5, 80°C | Thermococcus guaymasensis | |
1.1.1.2 | 49 | - |
meso-2,3-butanediol | cosubstrate: NADP+, pH 7.5, 80°C | Thermococcus guaymasensis | |
1.1.1.2 | 49 | - |
(2R,3R)-2,3-butanediol | cosubstrate: NADP+, pH 7.5, 80°C | Thermococcus guaymasensis | |
1.1.1.2 | 444 | - |
acetoin | cosubstrate: NADPH, pH 7.5, 80°C | Thermococcus guaymasensis | |
1.1.1.2 | 613 | - |
2-butanone | cosubstrate: NADPH, pH 7.5, 80°C | Thermococcus guaymasensis | |
1.1.1.2 | 962 | - |
diacetyl | cosubstrate: NADPH, pH 7.5, 80°C | Thermococcus guaymasensis | |
1.1.1.2 | 2192 | - |
2-butanol | cosubstrate: NADP+, pH 7.5, 80°C | Thermococcus guaymasensis | |
1.1.1.2 | 3333 | - |
NADP+ | cosubstrate: 2-butanol, pH 7.5, 80°C | Thermococcus guaymasensis | |
1.1.1.2 | 14360 | - |
NADPH | cosubstrate: 2-butanone, pH 7.5, 80°C | Thermococcus guaymasensis |