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Literature summary extracted from

  • Matsushima, A.; Sato, Y.; Otsuka, M.; Watanabe, T.; Yamamoto, H.; Hirata, T.
    An enone reductase from Nicotiana tabacum: cDNA cloning, expression in Escherichia coli, and reduction of enones with the recombinant proteins (2008), Bioorg. Chem., 36, 23-28.
    View publication on PubMed

Cloned(Commentary)

EC Number Cloned (Comment) 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 Comment 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] Comment 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 Comment Textmining
1.1.1.54 Nicotiana tabacum Q9SLN8
-
-
1.3.1.102 Nicotiana tabacum Q9SLN8
-
-

Purification (Commentary)

EC Number Purification (Comment) 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 Comment Substrates Organism Products Comment (Products) Rev. Reac.
1.1.1.54 (R)-pulegone + NADP+ recombinant enzyme 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 Nicotiana tabacum (1R,4R)-isomenthone + NADPH + H+
-
 ?
1.1.1.54 (S)-pulegone + NADP+ recombinant enzyme Nicotiana tabacum (1S,4S)-isomenthone + (1S,4R)-menthone + NADPH + H+
-
 ?
1.1.1.54 allyl alcohol + NADP+
-
 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 Nicotiana tabacum ?
-
 ?
1.3.1.102 (R)-pulegone + NADPH + H+
-
 Nicotiana tabacum (1R,4R)-isomenthone + (1R,4S)-menthone + NADP+
-
 ?
1.3.1.102 (S)-pulegone + NADPH + H+
-
 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 Nicotiana tabacum ?
-
 ?

Subunits

EC Number Subunits Comment 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 Comment 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] Comment 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 Comment 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 Comment Organism
1.1.1.54 7
-
 assay pH Nicotiana tabacum

Cofactor

EC Number Cofactor Comment Organism Structure
1.1.1.54 NADP+
-
 Nicotiana tabacum

kcat/KM [mM/s]

EC Number kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment 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