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Literature summary 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)

EC Number Cloned (Comment) Organism
1.1.1.206 gene TRI, sequence comparisons Dendrobium nobile
1.1.1.236 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

EC Number Protein Variants Comment Organism
1.1.1.236 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]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
1.1.1.236 29420
-
-
 Dendrobium nobile

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
1.1.1.206 tropinone + NADPH + H+ Dendrobium nobile
-
 tropine + NADP+
-
 r

Organism

EC Number Organism UniProt Comment Textmining
1.1.1.206 Dendrobium nobile H9BQR7
-
-
1.1.1.236 Dendrobium nobile H9BQR9
-
-

Purification (Commentary)

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

Source Tissue

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

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
1.1.1.206 3-quinuclidinone + NADPH + H+
-
 Dendrobium nobile 1-azabicyclo[2.2.2]octan-3-ol + NADP+
-
 r
1.1.1.206 4-methylcyclohexanone + NADPH + H+
-
 Dendrobium nobile 4-methylcyclohexanol + NADP+
-
 r
1.1.1.206 tropinone + NADPH + H+
-
 Dendrobium nobile tropine + NADP+
-
 r
1.1.1.236 3-quinuclidinone + NADPH + H+ wild-type and mutant Y201V enzymes Dendrobium nobile 1-azabicyclo[2.2.2]octan-3-ol + NADP+
-
 r
1.1.1.236 4-methylcyclohexanone + NADPH + H+ wild-type and mutant Y201V enzymes Dendrobium nobile 4-methylcyclohexanol + NADP+
-
 r
1.1.1.236 tropinone + NADPH + H+ recombinant enzyme mutant Y201V, no activity by wild-type TR2 enzyme Dendrobium nobile pseudotropine + NADP+
-
 r

Subunits

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

Synonyms

EC Number Synonyms Comment Organism
1.1.1.206 DnTR1
-
 Dendrobium nobile
1.1.1.206 TR1
-
 Dendrobium nobile
1.1.1.206 TRI
-
 Dendrobium nobile
1.1.1.236 DnTR2
-
 Dendrobium nobile
1.1.1.236 TR2
-
 Dendrobium nobile
1.1.1.236 TRII
-
 Dendrobium nobile

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
1.1.1.206 25
-
 assay at Dendrobium nobile
1.1.1.236 25
-
 assay at Dendrobium nobile

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
1.1.1.206 8
-
 tropinone reduction assay at Dendrobium nobile
1.1.1.236 8
-
 tropinone reduction assay at Dendrobium nobile

Cofactor

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

pI Value

EC Number Organism Comment pI Value Maximum pI Value
1.1.1.236 Dendrobium nobile sequence calculation
-
 6.44

Expression

EC Number Organism Comment Expression
1.1.1.206 Dendrobium nobile the enzyme expression is upregulated by methyl jasmonate, but not by salicylate and NO up
1.1.1.236 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

EC Number General Information Comment Organism
1.1.1.206 additional information three-dimensional enzyme structure comparisons, overview Dendrobium nobile
1.1.1.236 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
1.1.1.236 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
1.1.1.236 additional information three-dimensional structure modeling of DnTR2, catalytic triad Ser-Asn-Lys, role of Tyr201 in substrate binding, structure comparisons, overview Dendrobium nobile