BRENDA - Enzyme Database

An enone reductase from Nicotiana tabacum: cDNA cloning, expression in Escherichia coli, and reduction of enones with the recombinant proteins

Matsushima, A.; Sato, Y.; Otsuka, M.; Watanabe, T.; Yamamoto, H.; Hirata, T.; Bioorg. Chem. 36, 23-28 (2008) View publication on PubMed

Data extracted from this reference:

Cloned(Commentary)
EC Number
Cloned (Commentary)
Organism
1.1.1.54
GST-fusion protein expressed in Escherichia coli BL21
Nicotiana tabacum
1.3.1.102
NtRed-1 cloned in Escherichia coli
Nicotiana tabacum
KM Value [mM]
EC Number
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
1.1.1.54
0.025
-
(R)-pulegone
native enzyme, pH 7.0, 35°C
Nicotiana tabacum
1.1.1.54
1.4
-
(R)-pulegone
recombinant enzyme, pH 7.0, 35°C
Nicotiana tabacum
1.1.1.54
8.3
-
(S)-pulegone
recombinant enzyme, pH 7.0, 35°C
Nicotiana tabacum
1.3.1.102
1.4
-
(R)-pulegone
recombinant NtRed-1, pH 7.0 and 35°C
Nicotiana tabacum
1.3.1.102
8.3
-
(S)-pulegone
recombinant NtRed-1, pH 7.0 and 35°C
Nicotiana tabacum
Molecular Weight [Da]
EC Number
Molecular Weight [Da]
Molecular Weight Maximum [Da]
Commentary
Organism
1.1.1.54
36300
-
2 * 36300, SDS-PAGE, native mass by gel filtration
Nicotiana tabacum
1.1.1.54
38482
-
2 * 38482, recombinant protein, MALDI TOF-MS, native mass by gel filtration
Nicotiana tabacum
1.1.1.54
38487
-
2 * 38487, recombinant protein, calculated from the deduced amino acid sequence, native mass by gel filtration
Nicotiana tabacum
1.1.1.54
74000
-
gel filtration
Nicotiana tabacum
1.3.1.102
36300
-
native NtRed-1, SDS-PAGE
Nicotiana tabacum
1.3.1.102
38000
-
recombinant NtRed-1, 2 * 38000, gel filtration chromatography
Nicotiana tabacum
1.3.1.102
38480
-
recombinant NtRed-1, MALDI-TOF MS spectra
Nicotiana tabacum
1.3.1.102
74000
-
recombinant NtRed-1, gel filtration chromatography
Nicotiana tabacum
Organism
EC Number
Organism
UniProt
Commentary
Textmining
1.1.1.54
Nicotiana tabacum
Q9SLN8
-
-
1.3.1.102
Nicotiana tabacum
Q9SLN8
-
-
Purification (Commentary)
EC Number
Purification (Commentary)
Organism
1.1.1.54
native enzyme, recombinant enzyme purified using GST-tag
Nicotiana tabacum
1.3.1.102
by DEAE–Toyopearl and Red–Toyopearl column chromatography
Nicotiana tabacum
Substrates and Products (Substrate)
EC Number
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
Substrate Product ID
1.1.1.54
(R)-pulegone + NADP+
recombinant enzyme
685505
Nicotiana tabacum
(1R,4R)-isomenthone + (1R,4S)-menthone + NADPH + H+
reaction is not stereospecific as with the native enzyme
-
-
?
1.1.1.54
(R)-pulegone + NADP+
native enzyme
685505
Nicotiana tabacum
(1R,4R)-isomenthone + NADPH + H+
-
-
-
?
1.1.1.54
(S)-pulegone + NADP+
recombinant enzyme
685505
Nicotiana tabacum
(1S,4S)-isomenthone + (1S,4R)-menthone + NADPH + H+
-
-
-
?
1.1.1.54
allyl alcohol + NADP+
-
685505
Nicotiana tabacum
acrolein + NADPH + H+
-
-
-
?
1.1.1.54
additional information
(R)-carvone, (S)-carvone, (1S,5S)-verbenone, (1R,5R)-verbenone, oxalacetic acid, and (2S,4S)-carveol do not serve as substrate
685505
Nicotiana tabacum
?
-
-
-
?
1.3.1.102
(R)-pulegone + NADPH + H+
-
685505
Nicotiana tabacum
(1R,4R)-isomenthone + (1R,4S)-menthone + NADP+
-
-
-
?
1.3.1.102
(S)-pulegone + NADPH + H+
-
685505
Nicotiana tabacum
(1S,4S)-isomenthone + (1S,4R)-menthone + NADP+
-
-
-
?
1.3.1.102
additional information
recombinant NtRed-1 does not catalyze dehydrogenation of (2S,4S)-carveol, indicating that the recombinant NtRed-1 lacks the catalytic function as allyl-alcohol dehydrogenase, although the recombinant NtRed-1 is designed from the nucleotide sequence of allyl-ADH
685505
Nicotiana tabacum
?
-
-
-
?
Subunits
EC Number
Subunits
Commentary
Organism
1.1.1.54
dimer
2 * 36300, SDS-PAGE, native mass by gel filtration
Nicotiana tabacum
1.1.1.54
dimer
2 * 38482, recombinant protein, MALDI TOF-MS, native mass by gel filtration
Nicotiana tabacum
1.1.1.54
dimer
2 * 38487, recombinant protein, calculated from the deduced amino acid sequence, native mass by gel filtration
Nicotiana tabacum
1.3.1.102
homodimer
recombinant NtRed-1, 2 * 38000, gel filtration chromatography
Nicotiana tabacum
Synonyms
EC Number
Synonyms
Commentary
Organism
1.1.1.54
allyl-ADH
-
Nicotiana tabacum
1.1.1.54
NtRed-1
-
Nicotiana tabacum
1.3.1.102
allyl-alcohol dehydrogenase
-
Nicotiana tabacum
1.3.1.102
NtRed-1
-
Nicotiana tabacum
Temperature Optimum [°C]
EC Number
Temperature Optimum [°C]
Temperature Optimum Maximum [°C]
Commentary
Organism
1.1.1.54
35
-
assay temperature
Nicotiana tabacum
Turnover Number [1/s]
EC Number
Turnover Number Minimum [1/s]
Turnover Number Maximum [1/s]
Substrate
Commentary
Organism
Structure
1.1.1.54
2.8
-
(S)-pulegone
recombinant enzyme, pH 7.0, 35°C
Nicotiana tabacum
1.1.1.54
3.3
-
(R)-pulegone
recombinant enzyme, pH 7.0, 35°C
Nicotiana tabacum
1.3.1.102
2.8
-
(S)-pulegone
recombinant NtRed-1, pH 7.0 and 35°C
Nicotiana tabacum
1.3.1.102
3.3
-
(R)-pulegone
recombinant NtRed-1, pH 7.0 and 35°C
Nicotiana tabacum
pH Optimum
EC Number
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
1.1.1.54
7
-
assay pH
Nicotiana tabacum
Cofactor
EC Number
Cofactor
Commentary
Organism
Structure
1.1.1.54
NADP+
-
Nicotiana tabacum
Cloned(Commentary) (protein specific)
EC Number
Commentary
Organism
1.1.1.54
GST-fusion protein expressed in Escherichia coli BL21
Nicotiana tabacum
1.3.1.102
NtRed-1 cloned in Escherichia coli
Nicotiana tabacum
Cofactor (protein specific)
EC Number
Cofactor
Commentary
Organism
Structure
1.1.1.54
NADP+
-
Nicotiana tabacum
KM Value [mM] (protein specific)
EC Number
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
1.1.1.54
0.025
-
(R)-pulegone
native enzyme, pH 7.0, 35°C
Nicotiana tabacum
1.1.1.54
1.4
-
(R)-pulegone
recombinant enzyme, pH 7.0, 35°C
Nicotiana tabacum
1.1.1.54
8.3
-
(S)-pulegone
recombinant enzyme, pH 7.0, 35°C
Nicotiana tabacum
1.3.1.102
1.4
-
(R)-pulegone
recombinant NtRed-1, pH 7.0 and 35°C
Nicotiana tabacum
1.3.1.102
8.3
-
(S)-pulegone
recombinant NtRed-1, pH 7.0 and 35°C
Nicotiana tabacum
Molecular Weight [Da] (protein specific)
EC Number
Molecular Weight [Da]
Molecular Weight Maximum [Da]
Commentary
Organism
1.1.1.54
36300
-
2 * 36300, SDS-PAGE, native mass by gel filtration
Nicotiana tabacum
1.1.1.54
38482
-
2 * 38482, recombinant protein, MALDI TOF-MS, native mass by gel filtration
Nicotiana tabacum
1.1.1.54
38487
-
2 * 38487, recombinant protein, calculated from the deduced amino acid sequence, native mass by gel filtration
Nicotiana tabacum
1.1.1.54
74000
-
gel filtration
Nicotiana tabacum
1.3.1.102
36300
-
native NtRed-1, SDS-PAGE
Nicotiana tabacum
1.3.1.102
38000
-
recombinant NtRed-1, 2 * 38000, gel filtration chromatography
Nicotiana tabacum
1.3.1.102
38480
-
recombinant NtRed-1, MALDI-TOF MS spectra
Nicotiana tabacum
1.3.1.102
74000
-
recombinant NtRed-1, gel filtration chromatography
Nicotiana tabacum
Purification (Commentary) (protein specific)
EC Number
Commentary
Organism
1.1.1.54
native enzyme, recombinant enzyme purified using GST-tag
Nicotiana tabacum
1.3.1.102
by DEAE–Toyopearl and Red–Toyopearl column chromatography
Nicotiana tabacum
Substrates and Products (Substrate) (protein specific)
EC Number
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
ID
1.1.1.54
(R)-pulegone + NADP+
recombinant enzyme
685505
Nicotiana tabacum
(1R,4R)-isomenthone + (1R,4S)-menthone + NADPH + H+
reaction is not stereospecific as with the native enzyme
-
-
?
1.1.1.54
(R)-pulegone + NADP+
native enzyme
685505
Nicotiana tabacum
(1R,4R)-isomenthone + NADPH + H+
-
-
-
?
1.1.1.54
(S)-pulegone + NADP+
recombinant enzyme
685505
Nicotiana tabacum
(1S,4S)-isomenthone + (1S,4R)-menthone + NADPH + H+
-
-
-
?
1.1.1.54
allyl alcohol + NADP+
-
685505
Nicotiana tabacum
acrolein + NADPH + H+
-
-
-
?
1.1.1.54
additional information
(R)-carvone, (S)-carvone, (1S,5S)-verbenone, (1R,5R)-verbenone, oxalacetic acid, and (2S,4S)-carveol do not serve as substrate
685505
Nicotiana tabacum
?
-
-
-
?
1.3.1.102
(R)-pulegone + NADPH + H+
-
685505
Nicotiana tabacum
(1R,4R)-isomenthone + (1R,4S)-menthone + NADP+
-
-
-
?
1.3.1.102
(S)-pulegone + NADPH + H+
-
685505
Nicotiana tabacum
(1S,4S)-isomenthone + (1S,4R)-menthone + NADP+
-
-
-
?
1.3.1.102
additional information
recombinant NtRed-1 does not catalyze dehydrogenation of (2S,4S)-carveol, indicating that the recombinant NtRed-1 lacks the catalytic function as allyl-alcohol dehydrogenase, although the recombinant NtRed-1 is designed from the nucleotide sequence of allyl-ADH
685505
Nicotiana tabacum
?
-
-
-
?
Subunits (protein specific)
EC Number
Subunits
Commentary
Organism
1.1.1.54
dimer
2 * 36300, SDS-PAGE, native mass by gel filtration
Nicotiana tabacum
1.1.1.54
dimer
2 * 38482, recombinant protein, MALDI TOF-MS, native mass by gel filtration
Nicotiana tabacum
1.1.1.54
dimer
2 * 38487, recombinant protein, calculated from the deduced amino acid sequence, native mass by gel filtration
Nicotiana tabacum
1.3.1.102
homodimer
recombinant NtRed-1, 2 * 38000, gel filtration chromatography
Nicotiana tabacum
Temperature Optimum [°C] (protein specific)
EC Number
Temperature Optimum [°C]
Temperature Optimum Maximum [°C]
Commentary
Organism
1.1.1.54
35
-
assay temperature
Nicotiana tabacum
Turnover Number [1/s] (protein specific)
EC Number
Turnover Number Minimum [1/s]
Turnover Number Maximum [1/s]
Substrate
Commentary
Organism
Structure
1.1.1.54
2.8
-
(S)-pulegone
recombinant enzyme, pH 7.0, 35°C
Nicotiana tabacum
1.1.1.54
3.3
-
(R)-pulegone
recombinant enzyme, pH 7.0, 35°C
Nicotiana tabacum
1.3.1.102
2.8
-
(S)-pulegone
recombinant NtRed-1, pH 7.0 and 35°C
Nicotiana tabacum
1.3.1.102
3.3
-
(R)-pulegone
recombinant NtRed-1, pH 7.0 and 35°C
Nicotiana tabacum
pH Optimum (protein specific)
EC Number
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
1.1.1.54
7
-
assay pH
Nicotiana tabacum
KCat/KM [mM/s]
EC Number
kcat/KM Value [1/mMs-1]
kcat/KM Value Maximum [1/mMs-1]
Substrate
Commentary
Organism
Structure
1.3.1.102
0.34
-
(S)-pulegone
recombinant NtRed-1, pH 7.0 and 35°C
Nicotiana tabacum
1.3.1.102
2.4
-
(R)-pulegone
recombinant NtRed-1, pH 7.0 and 35°C
Nicotiana tabacum
KCat/KM [mM/s] (protein specific)
EC Number
KCat/KM Value [1/mMs-1]
KCat/KM Value Maximum [1/mMs-1]
Substrate
Commentary
Organism
Structure
1.3.1.102
0.34
-
(S)-pulegone
recombinant NtRed-1, pH 7.0 and 35°C
Nicotiana tabacum
1.3.1.102
2.4
-
(R)-pulegone
recombinant NtRed-1, pH 7.0 and 35°C
Nicotiana tabacum