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Literature summary for 1.1.1.184 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
Search for more literature for 1.1.1.184

Activating Compound

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

Application

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(Commentary)

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

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 [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

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

Molecular Weight [Da]

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 Stability

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

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 (Commentary)

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

Specific Activity [micromol/min/mg]

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 and Products (Substrate)

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

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

Synonyms

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

Temperature Range [°C]

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

Temperature Stability [°C]

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 [1/s]

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

pH Optimum Minimum pH Optimum Maximum Comment Organism
9
-
-
 Thermotoga maritima

Cofactor

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

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