Cloned (Comment) | Organism |
---|---|
gene DnTR1, sequence comparisons and unrooted phylogenetic tree, recombinant expression of His6-tagged enzyme in Escherichia coli strain BL21(DE3), realtime PCR enzyme expression analysis | Dendrobium nobile |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
additional information | - |
additional information | Michaelis-Menten kinetics | Dendrobium nobile | |
0.167 | - |
3-quinuclidinone | pH 8.0, 25°C, recombinant His-tagged enzyme | Dendrobium nobile | |
5.61 | - |
tropinone | pH 8.0, 25°C, recombinant His-tagged enzyme | Dendrobium nobile | |
8.71 | - |
4-methylcyclohexanone | pH 8.0, 25°C, recombinant His-tagged enzyme | Dendrobium nobile |
Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
---|---|---|---|
28000 | - |
- |
Dendrobium nobile |
60000 | - |
gel filtration, recombinant enzyme | Dendrobium nobile |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
tropinone + NADPH + H+ | Dendrobium nobile | - |
tropine + NADP+ | - |
r |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Dendrobium nobile | H9BQR7 | - |
- |
Purification (Comment) | Organism |
---|---|
recombinant His6-tagged enzyme from Escherichia coli strain BL21(DE3) by nickel affinity chromatography and gel filtration, followed by ultrafiltration | Dendrobium nobile |
Source Tissue | Comment | Organism | Textmining |
---|---|---|---|
leaf | - |
Dendrobium nobile | - |
additional information | DnTR1 may be ubiquitously expressed in all tissues, expression profile of DnTR1 | Dendrobium nobile | - |
root | - |
Dendrobium nobile | - |
stem | - |
Dendrobium nobile | - |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
3-quinuclidinone + NADPH + H+ | - |
Dendrobium nobile | 1-azabicyclo[2.2.2]octan-3-ol + NADP+ | - |
r | |
4-methylcyclohexanone + NADPH + H+ | - |
Dendrobium nobile | 4-methylcyclohexanol + NADP+ | - |
r | |
tropinone + NADPH + H+ | - |
Dendrobium nobile | tropine + NADP+ | - |
r |
Subunits | Comment | Organism |
---|---|---|
homodimer | 2 * 28000, recombinant enzyme, SDS-PAGE | Dendrobium nobile |
More | three-dimensional structure modelling of DnTR1, overview | Dendrobium nobile |
Synonyms | Comment | Organism |
---|---|---|
DnTR1 | - |
Dendrobium nobile |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
25 | - |
assay at | Dendrobium nobile |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
8 | - |
assay at | Dendrobium nobile |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
NADP+ | - |
Dendrobium nobile | |
NADPH | - |
Dendrobium nobile |
Organism | Comment | Expression |
---|---|---|
Dendrobium nobile | salicylic acid and sodium nitroprusside treatments do not affect the enzyme expression | additional information |
Dendrobium nobile | the expression of DnTR1 mRNA is enhanced 9.5times by treatment of methyl jasmonate after 24 h, expression profile of DnTR1 | up |
General Information | Comment | Organism |
---|---|---|
evolution | the enzyme belongs to the short chain dehydrogenase/reductase (SDR) superfamily that is composed of a group of NAD(P)H-dependent oxidoreductases that typically consist of 250-350 amino acids. Tropinone reductases (TRs) are a group of the SDR superfamily, that use the NADPH as coenzyme to reduce tropinone DnTR1 also contains the sequence pattern G-X3-G-X-G in the cofactor binding motif and active site motif (S-N-K) | Dendrobium nobile |
metabolism | tropinone reductases form a branch point in the pathway leading to tropine (TRI) and to pseudotropine (TRII) during the tropane alkaloid biosynthesis, overview. Enzyme TRI is involved in the formation of tropine, the preproduct of hyoscyamine, whereas enzyme TRII, EC 1.1.1.236, is responsible for the generation of pseudotropine | Dendrobium nobile |
additional information | three-dimensional structure modelling of DnTR1, overview. DnTR1 also contains the sequence pattern G-X3-G-X-G in the cofactor binding motif and active site motif (S-N-K). The positive and negative charges on the binding pockets surface of DsTRI are due to His112. The three catalytic residues are Ser158, Tyr171, and Lys175 in DsTRI | Dendrobium nobile |
physiological function | DnTR1 regulation may be involved in a jasmonate-dependent pathway | Dendrobium nobile |
kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.00094 | - |
tropinone | pH 8.0, 25°C, recombinant His-tagged enzyme | Dendrobium nobile | |
0.0032 | - |
4-methylcyclohexanone | pH 8.0, 25°C, recombinant His-tagged enzyme | Dendrobium nobile | |
22.18 | - |
3-quinuclidinone | pH 8.0, 25°C, recombinant His-tagged enzyme | Dendrobium nobile |