Activating Compound | Comment | Organism | Structure |
---|---|---|---|
EDTA | activtion to 112% activity | Thermotoga maritima | |
EDTA | up to 112% of initial activity | Thermotoga maritima |
Application | Comment | Organism |
---|---|---|
synthesis | optimum substrate, 2,2,2-trifluoroacetophenone, is asymmetrically reduced in a coupled NADPH-regeneration system with an enantioselectivity of 99.8% and a conversion of 98% | Thermotoga maritima |
Cloned (Comment) | Organism |
---|---|
expression in Escherichia coli | Thermotoga maritima |
gene Tm1743, expression of His-tagged enzyme in Escherichia coli strain BL21(DE3), subcloning in Escherichia coli strain DH5alpha | Thermotoga maritima |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
2-mercaptoethanol | 17% reduced activity; 87% residual activity | Thermotoga maritima | |
additional information | the nonionic surfactants Tween 80 and Triton X-100 have only weak impacts on the enzyme activity | Thermotoga maritima | |
Zn2+ | 1 mM, 97% inhibition; 97% inhibition at 1 mM | Thermotoga maritima |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.3 | - |
2,2,2-trifluoroacetophenone | pH 9.0, 70°C, recombinant enzyme | Thermotoga maritima | |
4.9 | - |
ethyl 2-methylacetoacetate | pH 9.0, 70°C, recombinant enzyme | Thermotoga maritima | |
6 | - |
ethyl 2-oxo-4-phenylbutyrate | pH 9.0, 70°C, recombinant enzyme | Thermotoga maritima |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
additional information | the enzyme activity is metal-ion-independent | Thermotoga maritima |
Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
---|---|---|---|
32000 | - |
x * 32000, recombinant His-tagged enzyme, SDS-PAGE | Thermotoga maritima |
32000 | - |
x * 32000, SDS-PAGE and calculated | Thermotoga maritima |
Organic Solvent | Comment | Organism |
---|---|---|
2-propanol | concentrations up to 10% v/v slightly increase the enzyme activity to 112% | Thermotoga maritima |
2-propanol | 10% v/v, 112% of initial activity | Thermotoga maritima |
acetonitrile | concentrations up to 10% v/v slightly increase the enzyme activity to 108% | Thermotoga maritima |
acetonitrile | 10% v/v, 108% of initial activity | Thermotoga maritima |
DMSO | concentrations up to 10% v/v slightly decrease the enzyme activity | Thermotoga maritima |
Ethanol | concentrations up to 10% v/v slightly increase the enzyme activity to 109% | Thermotoga maritima |
Ethanol | 10% v/v, 119% of initial activity | Thermotoga maritima |
Methanol | concentrations up to 10% v/v slightly decrease the enzyme activity | Thermotoga maritima |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Thermotoga maritima | Q9X265 | - |
- |
Thermotoga maritima | Q9X265 | gene Tm1743 | - |
Thermotoga maritima DSM 3109 | Q9X265 | - |
- |
Thermotoga maritima MSB8 / DSM 3109 / ATCC 43589 | Q9X265 | gene Tm1743 | - |
Purification (Comment) | Organism |
---|---|
recombinant His-tagged enzyme 17.8fold from Escherichia coli strain BL21(DE3) by affinity chromatography | Thermotoga maritima |
Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
---|---|---|---|
7.9 | - |
pH 9.0, 55°C | Thermotoga maritima |
7.9 | - |
purified recombinant His-tagged enzyme, pH 9.0, 70°C | Thermotoga maritima |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
2,2,2-trifluoroacetophenone + NADPH + H+ | the preferred substrate is asymmetrically reduced in a coupled NADPH-regeneration system with an enantioselectivity of 99.8% and a conversion of 98% | Thermotoga maritima | (1S)-2,2,2-trifluoro-1-phenylethanol + NADP+ | - |
r | |
2,2,2-trifluoroacetophenone + NADPH + H+ | best substrate, 462% of the activity with ethyl 2-methyl-3-oxobutanoate | Thermotoga maritima | (1S)-2,2,2-trifluoro-1-phenylethanol + NADP+ | - |
? | |
2,2,2-trifluoroacetophenone + NADPH + H+ | the preferred substrate is asymmetrically reduced in a coupled NADPH-regeneration system with an enantioselectivity of 99.8% and a conversion of 98% | Thermotoga maritima DSM 3109 | (1S)-2,2,2-trifluoro-1-phenylethanol + NADP+ | - |
r | |
2,2,2-trifluoroacetophenone + NADPH + H+ | the preferred substrate is asymmetrically reduced in a coupled NADPH-regeneration system with an enantioselectivity of 99.8% and a conversion of 98% | Thermotoga maritima MSB8 / DSM 3109 / ATCC 43589 | (1S)-2,2,2-trifluoro-1-phenylethanol + NADP+ | - |
r | |
benzaldehyde + NADPH + H+ | 43% of the activity with ethyl 2-methyl-3-oxobutanoate | Thermotoga maritima | benzyl alcohol + NADP+ | - |
? | |
benzaldehyde + NADPH + H+ | 43% of the activity with ethyl 2-methyl-3-oxobutanoate | Thermotoga maritima DSM 3109 | benzyl alcohol + NADP+ | - |
? | |
benzaldehyde + NADPH + H+ | 43% of the activity with ethyl 2-methyl-3-oxobutanoate | Thermotoga maritima MSB8 / DSM 3109 / ATCC 43589 | benzyl alcohol + NADP+ | - |
? | |
ethyl 2-methyl-3-oxobutanoate + NADPH + H+ | - |
Thermotoga maritima | ethyl 3-hydroxy-2-methylbutanoate + NADP+ | - |
? | |
ethyl 2-methylacetoacetate + NADPH + H+ | - |
Thermotoga maritima | ? + NADP+ | - |
? | |
ethyl 2-oxo-3-phenylpropanoate + NADPH + H+ | 128% of the activity with ethyl 2-methyl-3-oxobutanoate | Thermotoga maritima | ethyl 2-hydroxy-3-phenylpropanoate + NADP+ | - |
? | |
ethyl 2-oxo-4-phenylbutyrate + NADPH + H+ | - |
Thermotoga maritima | ? + NADP+ | - |
? | |
ethyl 2-oxopropanoate + NADPH + H+ | 41% of the activity with ethyl 2-methyl-3-oxobutanoate | Thermotoga maritima | ethyl 2-hydroxypropanoate + NADP+ | - |
? | |
ethyl 4-chloro-3-oxobutanoate + NADPH + H+ | 384% of the activity with ethyl 2-methyl-3-oxobutanoate | Thermotoga maritima | ethyl 4-chloro-3-hydroxybutanoate + NADP+ | - |
? | |
ethyl 4-chloro-3-oxobutanoate + NADPH + H+ | 384% of the activity with ethyl 2-methyl-3-oxobutanoate | Thermotoga maritima DSM 3109 | ethyl 4-chloro-3-hydroxybutanoate + NADP+ | - |
? | |
ethyl 4-chloro-3-oxobutanoate + NADPH + H+ | 384% of the activity with ethyl 2-methyl-3-oxobutanoate | Thermotoga maritima MSB8 / DSM 3109 / ATCC 43589 | ethyl 4-chloro-3-hydroxybutanoate + NADP+ | - |
? | |
additional information | the recombinant enzyme exhibits broad substrate specificity towards various keto esters, ketones and aldehydes, detailed overview | Thermotoga maritima | ? | - |
? | |
additional information | gene contains the conserved catalytic tetrad D-Y-K-H (Asp53, Tyr58, Lys84, and His117) and putative cofactor binding sites at Asp53, Ser147, Asn148, Gln169, Glu249, and Asn254 | Thermotoga maritima | ? | - |
? | |
additional information | the recombinant enzyme exhibits broad substrate specificity towards various keto esters, ketones and aldehydes, detailed overview | Thermotoga maritima DSM 3109 | ? | - |
? | |
additional information | gene contains the conserved catalytic tetrad D-Y-K-H (Asp53, Tyr58, Lys84, and His117) and putative cofactor binding sites at Asp53, Ser147, Asn148, Gln169, Glu249, and Asn254 | Thermotoga maritima DSM 3109 | ? | - |
? | |
additional information | the recombinant enzyme exhibits broad substrate specificity towards various keto esters, ketones and aldehydes, detailed overview | Thermotoga maritima MSB8 / DSM 3109 / ATCC 43589 | ? | - |
? | |
additional information | gene contains the conserved catalytic tetrad D-Y-K-H (Asp53, Tyr58, Lys84, and His117) and putative cofactor binding sites at Asp53, Ser147, Asn148, Gln169, Glu249, and Asn254 | Thermotoga maritima MSB8 / DSM 3109 / ATCC 43589 | ? | - |
? | |
propanal + NADPH + H+ | 54% of the activity with ethyl 2-methyl-3-oxobutanoate | Thermotoga maritima | propan-1-ol + NADP+ | - |
? |
Subunits | Comment | Organism |
---|---|---|
? | x * 32000, recombinant His-tagged enzyme, SDS-PAGE | Thermotoga maritima |
? | x * 32000, SDS-PAGE and calculated | Thermotoga maritima |
Synonyms | Comment | Organism |
---|---|---|
2,5-diketo-D-gluconic acid reductase | - |
Thermotoga maritima |
aldo-keto reductase | - |
Thermotoga maritima |
Tm1743 | - |
Thermotoga maritima |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
90 | - |
- |
Thermotoga maritima |
Temperature Minimum [°C] | Temperature Maximum [°C] | Comment | Organism |
---|---|---|---|
40 | 100 | activity range, profile overview | Thermotoga maritima |
Temperature Stability Minimum [°C] | Temperature Stability Maximum [°C] | Comment | Organism |
---|---|---|---|
30 | 60 | purified recombinant enzyme, stable for 15 h | Thermotoga maritima |
85 | - |
the purified recombinant enzyme retains 63% of its maximal activity after 15 h at 85°C | Thermotoga maritima |
85 | - |
15 h, 63% residual activity | Thermotoga maritima |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
11.2 | - |
ethyl 2-oxo-4-phenylbutyrate | pH 9.0, 70°C, recombinant enzyme | Thermotoga maritima | |
12.5 | - |
ethyl 2-methylacetoacetate | pH 9.0, 70°C, recombinant enzyme | Thermotoga maritima | |
16.4 | - |
2,2,2-trifluoroacetophenone | pH 9.0, 70°C, recombinant enzyme | Thermotoga maritima |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
9 | - |
- |
Thermotoga maritima |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
NADH | 42% of the activity with NADPH | Thermotoga maritima | |
NADP+ | - |
Thermotoga maritima | |
NADPH | dependent on | Thermotoga maritima | |
NADPH | preferred over NADH | Thermotoga maritima |
General Information | Comment | Organism |
---|---|---|
evolution | the enzyme belongs to the aldo-keto reductase (AKR) superfamily of NAD(P)(H)-dependent oxidoreductases | Thermotoga maritima |
additional information | conserved catalytic tetrad D-Y-K-H (Asp53, Tyr58, Lys84, and His117) and putative cofactor binding sites at Asp53, Ser147, Asn148, Gln169, Glu249, and Asn254 | Thermotoga maritima |