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Literature summary extracted from

  • Ma, Y.H.; Lv, D.Q.; Zhou, S.; Lai, D.Y.; Chen, Z.M.
    Characterization of an aldo-keto reductase from Thermotoga maritima with high thermostability and a broad substrate spectrum (2013), Biotechnol. Lett., 35, 757-762.
    View publication on PubMed

Activating Compound

EC Number Activating Compound Comment Organism Structure
1.1.1.184 EDTA activtion to 112% activity Thermotoga maritima
1.1.1.184 EDTA up to 112% of initial activity Thermotoga maritima

Application

EC Number Application Comment Organism
1.1.1.184 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(Commentary)

EC Number Cloned (Comment) Organism
1.1.1.184 expression in Escherichia coli Thermotoga maritima
1.1.1.184 gene Tm1743, expression of His-tagged enzyme in Escherichia coli strain BL21(DE3), subcloning in Escherichia coli strain DH5alpha Thermotoga maritima

Inhibitors

EC Number Inhibitors Comment Organism Structure
1.1.1.184 2-mercaptoethanol 17% reduced activity; 87% residual activity Thermotoga maritima
1.1.1.184 additional information the nonionic surfactants Tween 80 and Triton X-100 have only weak impacts on the enzyme activity Thermotoga maritima
1.1.1.184 Zn2+ 1 mM, 97% inhibition; 97% inhibition at 1 mM Thermotoga maritima

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
1.1.1.184 0.3
-
2,2,2-trifluoroacetophenone pH 9.0, 70°C, recombinant enzyme Thermotoga maritima
1.1.1.184 4.9
-
ethyl 2-methylacetoacetate pH 9.0, 70°C, recombinant enzyme Thermotoga maritima
1.1.1.184 6
-
ethyl 2-oxo-4-phenylbutyrate pH 9.0, 70°C, recombinant enzyme Thermotoga maritima

Metals/Ions

EC Number Metals/Ions Comment Organism Structure
1.1.1.184 additional information the enzyme activity is metal-ion-independent Thermotoga maritima

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
1.1.1.184 32000
-
x * 32000, recombinant His-tagged enzyme, SDS-PAGE Thermotoga maritima
1.1.1.184 32000
-
x * 32000, SDS-PAGE and calculated Thermotoga maritima

Organic Solvent Stability

EC Number Organic Solvent Comment Organism
1.1.1.184 2-propanol concentrations up to 10% v/v slightly increase the enzyme activity to 112% Thermotoga maritima
1.1.1.184 2-propanol 10% v/v, 112% of initial activity Thermotoga maritima
1.1.1.184 acetonitrile concentrations up to 10% v/v slightly increase the enzyme activity to 108% Thermotoga maritima
1.1.1.184 acetonitrile 10% v/v, 108% of initial activity Thermotoga maritima
1.1.1.184 DMSO concentrations up to 10% v/v slightly decrease the enzyme activity Thermotoga maritima
1.1.1.184 Ethanol concentrations up to 10% v/v slightly increase the enzyme activity to 109% Thermotoga maritima
1.1.1.184 Ethanol 10% v/v, 119% of initial activity Thermotoga maritima
1.1.1.184 Methanol concentrations up to 10% v/v slightly decrease the enzyme activity Thermotoga maritima

Organism

EC Number Organism UniProt Comment Textmining
1.1.1.184 Thermotoga maritima Q9X265
-
-
1.1.1.184 Thermotoga maritima Q9X265 gene Tm1743
-
1.1.1.184 Thermotoga maritima DSM 3109 Q9X265
-
-
1.1.1.184 Thermotoga maritima MSB8 / DSM 3109 / ATCC 43589 Q9X265 gene Tm1743
-

Purification (Commentary)

EC Number Purification (Comment) Organism
1.1.1.184 recombinant His-tagged enzyme 17.8fold from Escherichia coli strain BL21(DE3) by affinity chromatography Thermotoga maritima

Specific Activity [micromol/min/mg]

EC Number Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
1.1.1.184 7.9
-
pH 9.0, 55°C Thermotoga maritima
1.1.1.184 7.9
-
purified recombinant His-tagged enzyme, pH 9.0, 70°C Thermotoga maritima

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
1.1.1.184 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
1.1.1.184 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+
-
?
1.1.1.184 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
1.1.1.184 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
1.1.1.184 benzaldehyde + NADPH + H+ 43% of the activity with ethyl 2-methyl-3-oxobutanoate Thermotoga maritima benzyl alcohol + NADP+
-
?
1.1.1.184 benzaldehyde + NADPH + H+ 43% of the activity with ethyl 2-methyl-3-oxobutanoate Thermotoga maritima DSM 3109 benzyl alcohol + NADP+
-
?
1.1.1.184 benzaldehyde + NADPH + H+ 43% of the activity with ethyl 2-methyl-3-oxobutanoate Thermotoga maritima MSB8 / DSM 3109 / ATCC 43589 benzyl alcohol + NADP+
-
?
1.1.1.184 ethyl 2-methyl-3-oxobutanoate + NADPH + H+
-
Thermotoga maritima ethyl 3-hydroxy-2-methylbutanoate + NADP+
-
?
1.1.1.184 ethyl 2-methylacetoacetate + NADPH + H+
-
Thermotoga maritima ? + NADP+
-
?
1.1.1.184 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+
-
?
1.1.1.184 ethyl 2-oxo-4-phenylbutyrate + NADPH + H+
-
Thermotoga maritima ? + NADP+
-
?
1.1.1.184 ethyl 2-oxopropanoate + NADPH + H+ 41% of the activity with ethyl 2-methyl-3-oxobutanoate Thermotoga maritima ethyl 2-hydroxypropanoate + NADP+
-
?
1.1.1.184 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+
-
?
1.1.1.184 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+
-
?
1.1.1.184 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+
-
?
1.1.1.184 additional information the recombinant enzyme exhibits broad substrate specificity towards various keto esters, ketones and aldehydes, detailed overview Thermotoga maritima ?
-
?
1.1.1.184 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 ?
-
?
1.1.1.184 additional information the recombinant enzyme exhibits broad substrate specificity towards various keto esters, ketones and aldehydes, detailed overview Thermotoga maritima DSM 3109 ?
-
?
1.1.1.184 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 ?
-
?
1.1.1.184 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 ?
-
?
1.1.1.184 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 ?
-
?
1.1.1.184 propanal + NADPH + H+ 54% of the activity with ethyl 2-methyl-3-oxobutanoate Thermotoga maritima propan-1-ol + NADP+
-
?

Subunits

EC Number Subunits Comment Organism
1.1.1.184 ? x * 32000, recombinant His-tagged enzyme, SDS-PAGE Thermotoga maritima
1.1.1.184 ? x * 32000, SDS-PAGE and calculated Thermotoga maritima

Synonyms

EC Number Synonyms Comment Organism
1.1.1.184 2,5-diketo-D-gluconic acid reductase
-
Thermotoga maritima
1.1.1.184 aldo-keto reductase
-
Thermotoga maritima
1.1.1.184 Tm1743
-
Thermotoga maritima

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
1.1.1.184 90
-
-
Thermotoga maritima

Temperature Range [°C]

EC Number Temperature Minimum [°C] Temperature Maximum [°C] Comment Organism
1.1.1.184 40 100 activity range, profile overview Thermotoga maritima

Temperature Stability [°C]

EC Number Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
1.1.1.184 30 60 purified recombinant enzyme, stable for 15 h Thermotoga maritima
1.1.1.184 85
-
the purified recombinant enzyme retains 63% of its maximal activity after 15 h at 85°C Thermotoga maritima
1.1.1.184 85
-
15 h, 63% residual activity Thermotoga maritima

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
1.1.1.184 11.2
-
ethyl 2-oxo-4-phenylbutyrate pH 9.0, 70°C, recombinant enzyme Thermotoga maritima
1.1.1.184 12.5
-
ethyl 2-methylacetoacetate pH 9.0, 70°C, recombinant enzyme Thermotoga maritima
1.1.1.184 16.4
-
2,2,2-trifluoroacetophenone pH 9.0, 70°C, recombinant enzyme Thermotoga maritima

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
1.1.1.184 9
-
-
Thermotoga maritima

Cofactor

EC Number Cofactor Comment Organism Structure
1.1.1.184 NADH 42% of the activity with NADPH Thermotoga maritima
1.1.1.184 NADP+
-
Thermotoga maritima
1.1.1.184 NADPH dependent on Thermotoga maritima
1.1.1.184 NADPH preferred over NADH Thermotoga maritima

General Information

EC Number General Information Comment Organism
1.1.1.184 evolution the enzyme belongs to the aldo-keto reductase (AKR) superfamily of NAD(P)(H)-dependent oxidoreductases Thermotoga maritima
1.1.1.184 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