Cloned (Comment) | Organism |
---|---|
gene TRI, isolated from aerial tissue, DNA and amino acid sequence determination and analysis, sequence comparisons and phylogenetic analysis, overexpression of the His-tagged enzyme in Escherichia coli strain BL21 (DE3) | Withania coagulans |
gene WcTR-I, from leaves, DNA and amino acid sequence determination and analysis, sequence comparisons, phylogenetic analysis and tree, recombinant expression of His6-tagged enzyme in Escherichia coli strain BL21(DE3) | Withania coagulans |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
K+ | slight inhibitory effect at 5 mM | Withania coagulans | |
Li+ | slight inhibitory effect at 5 mM | Withania coagulans | |
Na+ | slight inhibitory effect at 5 mM | Withania coagulans | |
NH4+ | slight inhibitory effect at 5 mM | Withania coagulans |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
additional information | - |
additional information | the in vitro reaction kinetics predominantly favour the reaction direction of tropinone reduction, Michaelis-Menten kinetics | Withania coagulans | |
0.126 | - |
NADPH | pH 6.6, 30°C, recombinant enzyme | Withania coagulans | |
0.127 | - |
NADPH | pH 6.6, 30°C, recombinant His-tagged enzyme | Withania coagulans | |
0.208 | - |
NADP+ | pH 6.6, 30°C, recombinant enzyme | Withania coagulans | |
0.208 | - |
NADP+ | pH 6.6, 30°C, recombinant His-tagged enzyme | Withania coagulans | |
1.451 | - |
tropinone | pH 6.6, 30°C, recombinant enzyme | Withania coagulans | |
1.451 | - |
tropinone | pH 6.6, 30°C, recombinant His-tagged enzyme | Withania coagulans | |
26.31 | - |
Tropine | pH 6.6, 30°C, recombinant enzyme | Withania coagulans | |
26.31 | - |
Tropine | pH 6.6, 30°C, recombinant His-tagged enzyme | Withania coagulans |
Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
---|---|---|---|
29340 | - |
x * 29340, sequence calculation | Withania coagulans |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
additional information | Withania coagulans | the enzyme TR-I has a wide substrate specificity but does not cover the substrates of other well-known plant SDR related to menthol metabolism | ? | - |
? | |
tropinone + NADPH + H+ | Withania coagulans | preferred substrate | tropine + NADP+ | - |
r | |
tropinone + NADPH + H+ | Withania coagulans | the reaction direction of tropinone reduction is predominantly operational in planta | tropine + NADP+ | - |
r |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Withania coagulans | L7QI79 | - |
- |
Purification (Comment) | Organism |
---|---|
recombinant His-tagged enzyme from Escherichia coli strain BL21 (DE3) by nickel affinity chromatography | Withania coagulans |
recombinant His6-tagged enzyme from Escherichia coli strain BL21(DE3) by nickel affinity chromatography | Withania coagulans |
Source Tissue | Comment | Organism | Textmining |
---|---|---|---|
leaf | - |
Withania coagulans | - |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
1-methylpyrrolidine + NADPH + H+ | - |
Withania coagulans | ? + NADP+ | - |
? | |
1-propyl-4-piperidone + NADPH + H+ | - |
Withania coagulans | ? + NADP+ | - |
? | |
2-pyrrolidone + NADPH + H+ | - |
Withania coagulans | ? + NADP+ | - |
? | |
3-methylcyclohexanone + NADPH + H+ | - |
Withania coagulans | ? + NADP+ | - |
? | |
3-methylcyclohexanone + NADPH + H+ | - |
Withania coagulans | 3-methylcyclohexanol + NADP+ | - |
? | |
4-chlor-1-methylpiperidine + NADPH + H+ | - |
Withania coagulans | ? + NADP+ | - |
? | |
4-ethylcyclohexanone + NADPH + H+ | - |
Withania coagulans | 4-ethylcyclohexanol + NADP+ | - |
? | |
4-methylcyclohexanone + NADPH + H+ | - |
Withania coagulans | 4-methylcyclohexanol + NADP+ | - |
? | |
4-piperidone + NADPH + H+ | - |
Withania coagulans | ? + NADP+ | - |
? | |
exo-6-hydroxytropinone + NADPH + H+ | - |
Withania coagulans | ? + NADP+ | - |
? | |
additional information | the enzyme TR-I has a wide substrate specificity but does not cover the substrates of other well-known plant SDR related to menthol metabolism | Withania coagulans | ? | - |
? | |
additional information | no activity by TR-I with 1-methyl-2-pyrrolidone | Withania coagulans | ? | - |
? | |
additional information | wide substrate specificity, overview. No activity with 1-methyl-2-pyrrolidone. The reverse reaction of tropine oxidation is highly specific and none of the tested alcohols can be oxidized with either NADP+ or NAD+ as cofactor | Withania coagulans | ? | - |
? | |
N-methyl-4-piperidone + NADPH + H+ | - |
Withania coagulans | ? + NADP+ | - |
? | |
N-methylpiperidine + NADPH + H+ | - |
Withania coagulans | ? + NADP+ | - |
? | |
tetrahydro-4H-thiopyran-4-one + NADPH + H+ | - |
Withania coagulans | ? + NADP+ | - |
? | |
tropine + NADP+ | in vitro reaction kinetics show that the enzyme predominantly favours the reverse reaction | Withania coagulans | tropinone + NADPH + H+ | - |
r | |
tropinone + NADPH + H+ | best substrate | Withania coagulans | tropine + NADP+ | - |
r | |
tropinone + NADPH + H+ | preferred substrate | Withania coagulans | tropine + NADP+ | - |
r | |
tropinone + NADPH + H+ | the reaction direction of tropinone reduction is predominantly operational in planta | Withania coagulans | tropine + NADP+ | - |
r | |
tropinone + NADPH + H+ | the in vitro reaction kinetics predominantly favour the reaction direction of tropinone reduction | Withania coagulans | tropine + NADP+ | - |
r |
Subunits | Comment | Organism |
---|---|---|
? | x * 29340, sequence calculation | Withania coagulans |
More | superimposition of three-dimensional models of tropinone reductases,overview | Withania coagulans |
More | a three dimensional model is prepared by taking Datura stramonium TR-II, PDB ID 1ipf, as template, structure comparison, overview | Withania coagulans |
Synonyms | Comment | Organism |
---|---|---|
TR-I | - |
Withania coagulans |
tropine biosynthetic tropinone reductase | - |
Withania coagulans |
tropinone reductase I | - |
Withania coagulans |
WcTR-I | - |
Withania coagulans |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
30 | - |
assay at | Withania coagulans |
Temperature Stability Minimum [°C] | Temperature Stability Maximum [°C] | Comment | Organism |
---|---|---|---|
50 | - |
purified recombinant enzyme, 30 min, over 92% of maximal activity remaining | Withania coagulans |
50 | 80 | purified recombinant His-tagged enzyme, pH 6.6, 30 min, over 92% activity remaining at up to 50°C, 30% remaining at 60°C, 7% at 70°C, complete inactivation occurs at 80°C | Withania coagulans |
60 | - |
purified recombinant enzyme, 30 min, 30% of maximal activity remaining | Withania coagulans |
70 | - |
purified recombinant enzyme, 30 min, 7% of maximal activity remaining | Withania coagulans |
80 | - |
purified recombinant enzyme, 30 min, inactivation | Withania coagulans |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
1.87 | - |
Tropine | pH 6.6, 30°C, recombinant enzyme | Withania coagulans | |
1.87 | - |
Tropine | pH 6.6, 30°C, recombinant His-tagged enzyme | Withania coagulans | |
2.94 | - |
NADP+ | pH 6.6, 30°C, recombinant enzyme | Withania coagulans | |
2.94 | - |
NADP+ | pH 6.6, 30°C, recombinant His-tagged enzyme | Withania coagulans | |
6.76 | - |
NADPH | pH 6.6, 30°C, recombinant enzyme | Withania coagulans | |
6.76 | - |
NADPH | pH 6.6, 30°C, recombinant His-tagged enzyme | Withania coagulans | |
16.74 | - |
tropinone | pH 6.6, 30°C, recombinant enzyme | Withania coagulans | |
16.74 | - |
tropinone | pH 6.6, 30°C, recombinant His-tagged enzyme | Withania coagulans |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
6.6 | - |
- |
Withania coagulans |
6.6 | - |
tropinone reduction | Withania coagulans |
pH Minimum | pH Maximum | Comment | Organism |
---|---|---|---|
additional information | - |
narrow pH range in tropinone reduction | Withania coagulans |
6.1 | 7 | narrow pH range | Withania coagulans |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
additional information | the enzyme has absolute specificity for NADPH/NADP+. The affinity of the enzyme for NADPH is much higher than for NADP+ compared to NADPH | Withania coagulans | |
NADP+ | - |
Withania coagulans | |
NADPH | - |
Withania coagulans |
Organism | Comment | pI Value Maximum | pI Value |
---|---|---|---|
Withania coagulans | sequence calculation | - |
6.12 |
General Information | Comment | Organism |
---|---|---|
evolution | tropinone reductase TR-I belongs to the short chain dehydrogenase/reductase family, SDR, of enzymes and has a YSASK as the signature YXXXK motif of SDRs, catalytic tetrade comprises N, S, Y, and K residues, overview | Withania coagulans |
evolution | tropinone reductases (TRs) are small proteins belonging to the SDR (short chain dehydrogenase/reductase) family of enzymes. The enzyme's sequence contains the signature YXXXK motif of SDRs. Residues His112, Ala160, Val168, Ile223 and Phe226 are conserved in the TRs, and conserved at corresponding positions in WcTR-I, and involved in stereospecificity of the respective TR-Is. Tropane alkaloids might have evolved independently in plants, at least in Solanaceae and Erythroxylaceae | Withania coagulans |
metabolism | the enzyme catalyzes a step intermediary enroute to biosynthesis of tropane esters of medicinal importance, i.e. hyoscyamine/scopolamine | Withania coagulans |
metabolism | enzymes TR-I and TR-II catalyze the conversion of tropinone into tropane alcohols (tropine and pseudotropine, respectively). The steps are intermediary enroute to biosynthesis of tropane esters of medicinal importance, hyoscyamine/scopolamine, and calystegins, respectively. The enzyme has a wide substrate specificity but does not cover the substrates of other plant SDR enzymes related to menthol metabolism. The leaf tissue expression of the enzyme with these catalytic features suggests physiologically much prodigal rate of operation of the tropane alkaloid pathway in the leaf tissue itself | Withania coagulans |
additional information | superimposition of three-dimensional models of tropinone reductases,overview | Withania coagulans |
additional information | a three dimensional model is prepared by taking Datura stramonium TR-II, PDB ID 1ipf, as template, structure comparison, overview | Withania coagulans |
kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.071 | - |
Tropine | pH 6.6, 30°C, recombinant enzyme | Withania coagulans | |
0.071 | - |
Tropine | pH 6.6, 30°C, recombinant His-tagged enzyme | Withania coagulans | |
11.53 | - |
tropinone | pH 6.6, 30°C, recombinant enzyme | Withania coagulans | |
11.53 | - |
tropinone | pH 6.6, 30°C, recombinant His-tagged enzyme | Withania coagulans | |
14 | - |
NADP+ | pH 6.6, 30°C, recombinant enzyme | Withania coagulans | |
14 | - |
NADP+ | pH 6.6, 30°C, recombinant His-tagged enzyme | Withania coagulans | |
53 | - |
NADPH | pH 6.6, 30°C, recombinant enzyme | Withania coagulans | |
53 | - |
NADPH | pH 6.6, 30°C, recombinant His-tagged enzyme | Withania coagulans |