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Literature summary for 1.1.1.236 extracted from

  • Cheng, X.; Chen, W.; Zhou, Z.; Liu, J.; Wang, H.
    Functional characterization of a novel tropinone reductase-like gene in Dendrobium nobile Lindl (2013), J. Plant Physiol., 170, 958-964.
    View publication on PubMed

Cloned(Commentary)

Cloned (Comment) Organism
gene TRII, DNA and amino acid sequence determination and analysis, phylogenetic analysis and tree, recombinant expression of His6-tagged wild-type and mutant enzymes in Escherichia coli strain BL21(DE3), realtime PCR enzyme expression analysis Dendrobium nobile

Protein Variants

Protein Variants Comment Organism
Y201V site-directed mutagenesis, the mutant enzyme shows restored activity with tropinone, while the wild-type is inactive with tropinone. The mutant enzymes also remain to have the ability to reduce the 3-quinuclidinone and 4-methylcyclohexanone, although the Vmax of DnTR2-Y201V toward 4-methylcyclohexanone and 3-quinuclidinone are significantly lower (23 and 3fold decrease, respectively) than that of wild-type DnTR2 protein Dendrobium nobile

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
29420
-
-
Dendrobium nobile

Organism

Organism UniProt Comment Textmining
Dendrobium nobile H9BQR9
-
-

Purification (Commentary)

Purification (Comment) Organism
recombinant His6-tagged wild-type and mutant enzymes from Escherichia coli strain BL21(DE3) by nickel affinity chromatography Dendrobium nobile

Source Tissue

Source Tissue Comment Organism Textmining
leaf
-
Dendrobium nobile
-
additional information TR2 shows constitutive expression in all tissues Dendrobium nobile
-
root
-
Dendrobium nobile
-
stem
-
Dendrobium nobile
-

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
3-quinuclidinone + NADPH + H+ wild-type and mutant Y201V enzymes Dendrobium nobile 1-azabicyclo[2.2.2]octan-3-ol + NADP+
-
r
4-methylcyclohexanone + NADPH + H+ wild-type and mutant Y201V enzymes Dendrobium nobile 4-methylcyclohexanol + NADP+
-
r
tropinone + NADPH + H+ recombinant enzyme mutant Y201V, no activity by wild-type TR2 enzyme Dendrobium nobile pseudotropine + NADP+
-
r

Subunits

Subunits Comment Organism
? x * 29420, sequence calculation Dendrobium nobile
More three-dimensional structure modeling of DnTR2, structure comparisons, overview Dendrobium nobile

Synonyms

Synonyms Comment Organism
DnTR2
-
Dendrobium nobile
TR2
-
Dendrobium nobile
TRII
-
Dendrobium nobile

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
25
-
assay at Dendrobium nobile

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
8
-
tropinone reduction assay at Dendrobium nobile

Cofactor

Cofactor Comment Organism Structure
additional information the enzyme contains the conserved NAD(P)H binding motif (Gly-X3-Gly-X-Gly) Dendrobium nobile
NADP+
-
Dendrobium nobile
NADPH
-
Dendrobium nobile

pI Value

Organism Comment pI Value Maximum pI Value
Dendrobium nobile sequence calculation
-
6.44

Expression

Organism Comment Expression
Dendrobium nobile the enzyme expression is induced in leaves, stems, and rootsby methyljasmonate (highest in leaves, peaks after 24 h), and salicylate (peaks after 8 h), as well as by NO (peaks after 16 h) up

General Information

General Information Comment Organism
evolution DnTR2 amino acid sequence contains a conserved Rossmann folding structure, which includes a conserved NAD(P)H binding motif (Gly-X3-Gly-X-Gly) and catalytic residues Ser-Asn-Lys, suggesting that DnTR2 is a member of the SDR superfamily Dendrobium nobile
malfunction DnTR2 residue Tyr201 is located at the opposite side of Arg110 in the inner substrate binding surface. These structural characters suggest that the disabled tropinone reduction activity of DnTR2 may be caused by the replacement of an uncharged amino acid at position 201 Dendrobium nobile
additional information three-dimensional structure modeling of DnTR2, catalytic triad Ser-Asn-Lys, role of Tyr201 in substrate binding, structure comparisons, overview Dendrobium nobile