BRENDA - Enzyme Database

Molecular cloning and catalytic characterization of a recombinant tropine biosynthetic tropinone reductase from Withania coagulans leaf

Kushwaha, A.K.; Sangwan, N.S.; Tripathi, S.; Sangwan, R.S.; Gene 516, 238-247 (2013) View publication on PubMed

Data extracted from this reference:

Cloned(Commentary)
EC Number
Cloned (Commentary)
Organism
1.1.1.206
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
1.1.1.206
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
EC Number
Inhibitors
Commentary
Organism
Structure
1.1.1.206
K+
slight inhibitory effect at 5 mM
Withania coagulans
1.1.1.206
Li+
slight inhibitory effect at 5 mM
Withania coagulans
1.1.1.206
Na+
slight inhibitory effect at 5 mM
Withania coagulans
1.1.1.206
NH4+
slight inhibitory effect at 5 mM
Withania coagulans
KM Value [mM]
EC Number
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
1.1.1.206
additional information
-
additional information
the in vitro reaction kinetics predominantly favour the reaction direction of tropinone reduction, Michaelis-Menten kinetics
Withania coagulans
1.1.1.206
0.126
-
NADPH
pH 6.6, 30°C, recombinant enzyme
Withania coagulans
1.1.1.206
0.127
-
NADPH
pH 6.6, 30°C, recombinant His-tagged enzyme
Withania coagulans
1.1.1.206
0.208
-
NADP+
pH 6.6, 30°C, recombinant enzyme
Withania coagulans
1.1.1.206
0.208
-
NADP+
pH 6.6, 30°C, recombinant His-tagged enzyme
Withania coagulans
1.1.1.206
1.451
-
tropinone
pH 6.6, 30°C, recombinant enzyme
Withania coagulans
1.1.1.206
1.451
-
tropinone
pH 6.6, 30°C, recombinant His-tagged enzyme
Withania coagulans
1.1.1.206
26.31
-
Tropine
pH 6.6, 30°C, recombinant enzyme
Withania coagulans
1.1.1.206
26.31
-
Tropine
pH 6.6, 30°C, recombinant His-tagged enzyme
Withania coagulans
Molecular Weight [Da]
EC Number
Molecular Weight [Da]
Molecular Weight Maximum [Da]
Commentary
Organism
1.1.1.206
29340
-
x * 29340, sequence calculation
Withania coagulans
Natural Substrates/ Products (Substrates)
EC Number
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
ID
1.1.1.206
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
?
-
-
?
1.1.1.206
tropinone + NADPH + H+
Withania coagulans
preferred substrate
tropine + NADP+
-
-
r
1.1.1.206
tropinone + NADPH + H+
Withania coagulans
the reaction direction of tropinone reduction is predominantly operational in planta
tropine + NADP+
-
-
r
Organism
EC Number
Organism
UniProt
Commentary
Textmining
1.1.1.206
Withania coagulans
L7QI79
-
-
Purification (Commentary)
EC Number
Purification (Commentary)
Organism
1.1.1.206
recombinant His-tagged enzyme from Escherichia coli strain BL21 (DE3) by nickel affinity chromatography
Withania coagulans
1.1.1.206
recombinant His6-tagged enzyme from Escherichia coli strain BL21(DE3) by nickel affinity chromatography
Withania coagulans
Source Tissue
EC Number
Source Tissue
Commentary
Organism
Textmining
1.1.1.206
leaf
-
Withania coagulans
-
Substrates and Products (Substrate)
EC Number
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
Substrate Product ID
1.1.1.206
1-methylpyrrolidine + NADPH + H+
-
725098
Withania coagulans
? + NADP+
-
-
-
?
1.1.1.206
1-propyl-4-piperidone + NADPH + H+
-
725098
Withania coagulans
? + NADP+
-
-
-
?
1.1.1.206
2-pyrrolidone + NADPH + H+
-
725098
Withania coagulans
? + NADP+
-
-
-
?
1.1.1.206
3-methylcyclohexanone + NADPH + H+
-
725098
Withania coagulans
? + NADP+
-
-
-
?
1.1.1.206
3-methylcyclohexanone + NADPH + H+
-
725098
Withania coagulans
3-methylcyclohexanol + NADP+
-
-
-
?
1.1.1.206
4-chlor-1-methylpiperidine + NADPH + H+
-
725098
Withania coagulans
? + NADP+
-
-
-
?
1.1.1.206
4-ethylcyclohexanone + NADPH + H+
-
725098
Withania coagulans
4-ethylcyclohexanol + NADP+
-
-
-
?
1.1.1.206
4-methylcyclohexanone + NADPH + H+
-
725098
Withania coagulans
4-methylcyclohexanol + NADP+
-
-
-
?
1.1.1.206
4-piperidone + NADPH + H+
-
725098
Withania coagulans
? + NADP+
-
-
-
?
1.1.1.206
exo-6-hydroxytropinone + NADPH + H+
-
725098
Withania coagulans
? + NADP+
-
-
-
?
1.1.1.206
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
725098
Withania coagulans
?
-
-
-
?
1.1.1.206
additional information
no activity by TR-I with 1-methyl-2-pyrrolidone
725098
Withania coagulans
?
-
-
-
?
1.1.1.206
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
725098
Withania coagulans
?
-
-
-
?
1.1.1.206
N-methyl-4-piperidone + NADPH + H+
-
725098
Withania coagulans
? + NADP+
-
-
-
?
1.1.1.206
N-methylpiperidine + NADPH + H+
-
725098
Withania coagulans
? + NADP+
-
-
-
?
1.1.1.206
tetrahydro-4H-thiopyran-4-one + NADPH + H+
-
725098
Withania coagulans
? + NADP+
-
-
-
?
1.1.1.206
tropine + NADP+
in vitro reaction kinetics show that the enzyme predominantly favours the reverse reaction
725098
Withania coagulans
tropinone + NADPH + H+
-
-
-
r
1.1.1.206
tropinone + NADPH + H+
best substrate
725098
Withania coagulans
tropine + NADP+
-
-
-
r
1.1.1.206
tropinone + NADPH + H+
preferred substrate
725098
Withania coagulans
tropine + NADP+
-
-
-
r
1.1.1.206
tropinone + NADPH + H+
the reaction direction of tropinone reduction is predominantly operational in planta
725098
Withania coagulans
tropine + NADP+
-
-
-
r
1.1.1.206
tropinone + NADPH + H+
the in vitro reaction kinetics predominantly favour the reaction direction of tropinone reduction
725098
Withania coagulans
tropine + NADP+
-
-
-
r
Subunits
EC Number
Subunits
Commentary
Organism
1.1.1.206
?
x * 29340, sequence calculation
Withania coagulans
1.1.1.206
More
superimposition of three-dimensional models of tropinone reductases,overview
Withania coagulans
1.1.1.206
More
a three dimensional model is prepared by taking Datura stramonium TR-II, PDB ID 1ipf, as template, structure comparison, overview
Withania coagulans
Synonyms
EC Number
Synonyms
Commentary
Organism
1.1.1.206
TR-I
-
Withania coagulans
1.1.1.206
tropine biosynthetic tropinone reductase
-
Withania coagulans
1.1.1.206
tropinone reductase I
-
Withania coagulans
1.1.1.206
WcTR-I
-
Withania coagulans
Temperature Optimum [°C]
EC Number
Temperature Optimum [°C]
Temperature Optimum Maximum [°C]
Commentary
Organism
1.1.1.206
30
-
assay at
Withania coagulans
Temperature Stability [°C]
EC Number
Temperature Stability Minimum [°C]
Temperature Stability Maximum [°C]
Commentary
Organism
1.1.1.206
50
-
purified recombinant enzyme, 30 min, over 92% of maximal activity remaining
Withania coagulans
1.1.1.206
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
1.1.1.206
60
-
purified recombinant enzyme, 30 min, 30% of maximal activity remaining
Withania coagulans
1.1.1.206
70
-
purified recombinant enzyme, 30 min, 7% of maximal activity remaining
Withania coagulans
1.1.1.206
80
-
purified recombinant enzyme, 30 min, inactivation
Withania coagulans
Turnover Number [1/s]
EC Number
Turnover Number Minimum [1/s]
Turnover Number Maximum [1/s]
Substrate
Commentary
Organism
Structure
1.1.1.206
1.87
-
Tropine
pH 6.6, 30°C, recombinant enzyme
Withania coagulans
1.1.1.206
1.87
-
Tropine
pH 6.6, 30°C, recombinant His-tagged enzyme
Withania coagulans
1.1.1.206
2.94
-
NADP+
pH 6.6, 30°C, recombinant enzyme
Withania coagulans
1.1.1.206
2.94
-
NADP+
pH 6.6, 30°C, recombinant His-tagged enzyme
Withania coagulans
1.1.1.206
6.76
-
NADPH
pH 6.6, 30°C, recombinant enzyme
Withania coagulans
1.1.1.206
6.76
-
NADPH
pH 6.6, 30°C, recombinant His-tagged enzyme
Withania coagulans
1.1.1.206
16.74
-
tropinone
pH 6.6, 30°C, recombinant enzyme
Withania coagulans
1.1.1.206
16.74
-
tropinone
pH 6.6, 30°C, recombinant His-tagged enzyme
Withania coagulans
pH Optimum
EC Number
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
1.1.1.206
6.6
-
-
Withania coagulans
1.1.1.206
6.6
-
tropinone reduction
Withania coagulans
pH Range
EC Number
pH Minimum
pH Maximum
Commentary
Organism
1.1.1.206
additional information
-
narrow pH range in tropinone reduction
Withania coagulans
1.1.1.206
6.1
7
narrow pH range
Withania coagulans
Cofactor
EC Number
Cofactor
Commentary
Organism
Structure
1.1.1.206
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
1.1.1.206
NADP+
-
Withania coagulans
1.1.1.206
NADPH
-
Withania coagulans
pI Value
EC Number
Organism
Commentary
pI Value Maximum
pI Value
1.1.1.206
Withania coagulans
sequence calculation
-
6.12
Cloned(Commentary) (protein specific)
EC Number
Commentary
Organism
1.1.1.206
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
1.1.1.206
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
Cofactor (protein specific)
EC Number
Cofactor
Commentary
Organism
Structure
1.1.1.206
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
1.1.1.206
NADP+
-
Withania coagulans
1.1.1.206
NADPH
-
Withania coagulans
Inhibitors (protein specific)
EC Number
Inhibitors
Commentary
Organism
Structure
1.1.1.206
K+
slight inhibitory effect at 5 mM
Withania coagulans
1.1.1.206
Li+
slight inhibitory effect at 5 mM
Withania coagulans
1.1.1.206
Na+
slight inhibitory effect at 5 mM
Withania coagulans
1.1.1.206
NH4+
slight inhibitory effect at 5 mM
Withania coagulans
KM Value [mM] (protein specific)
EC Number
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
1.1.1.206
additional information
-
additional information
the in vitro reaction kinetics predominantly favour the reaction direction of tropinone reduction, Michaelis-Menten kinetics
Withania coagulans
1.1.1.206
0.126
-
NADPH
pH 6.6, 30°C, recombinant enzyme
Withania coagulans
1.1.1.206
0.127
-
NADPH
pH 6.6, 30°C, recombinant His-tagged enzyme
Withania coagulans
1.1.1.206
0.208
-
NADP+
pH 6.6, 30°C, recombinant enzyme
Withania coagulans
1.1.1.206
0.208
-
NADP+
pH 6.6, 30°C, recombinant His-tagged enzyme
Withania coagulans
1.1.1.206
1.451
-
tropinone
pH 6.6, 30°C, recombinant enzyme
Withania coagulans
1.1.1.206
1.451
-
tropinone
pH 6.6, 30°C, recombinant His-tagged enzyme
Withania coagulans
1.1.1.206
26.31
-
Tropine
pH 6.6, 30°C, recombinant enzyme
Withania coagulans
1.1.1.206
26.31
-
Tropine
pH 6.6, 30°C, recombinant His-tagged enzyme
Withania coagulans
Molecular Weight [Da] (protein specific)
EC Number
Molecular Weight [Da]
Molecular Weight Maximum [Da]
Commentary
Organism
1.1.1.206
29340
-
x * 29340, sequence calculation
Withania coagulans
Natural Substrates/ Products (Substrates) (protein specific)
EC Number
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
ID
1.1.1.206
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
?
-
-
?
1.1.1.206
tropinone + NADPH + H+
Withania coagulans
preferred substrate
tropine + NADP+
-
-
r
1.1.1.206
tropinone + NADPH + H+
Withania coagulans
the reaction direction of tropinone reduction is predominantly operational in planta
tropine + NADP+
-
-
r
Purification (Commentary) (protein specific)
EC Number
Commentary
Organism
1.1.1.206
recombinant His-tagged enzyme from Escherichia coli strain BL21 (DE3) by nickel affinity chromatography
Withania coagulans
1.1.1.206
recombinant His6-tagged enzyme from Escherichia coli strain BL21(DE3) by nickel affinity chromatography
Withania coagulans
Source Tissue (protein specific)
EC Number
Source Tissue
Commentary
Organism
Textmining
1.1.1.206
leaf
-
Withania coagulans
-
Substrates and Products (Substrate) (protein specific)
EC Number
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
ID
1.1.1.206
1-methylpyrrolidine + NADPH + H+
-
725098
Withania coagulans
? + NADP+
-
-
-
?
1.1.1.206
1-propyl-4-piperidone + NADPH + H+
-
725098
Withania coagulans
? + NADP+
-
-
-
?
1.1.1.206
2-pyrrolidone + NADPH + H+
-
725098
Withania coagulans
? + NADP+
-
-
-
?
1.1.1.206
3-methylcyclohexanone + NADPH + H+
-
725098
Withania coagulans
? + NADP+
-
-
-
?
1.1.1.206
3-methylcyclohexanone + NADPH + H+
-
725098
Withania coagulans
3-methylcyclohexanol + NADP+
-
-
-
?
1.1.1.206
4-chlor-1-methylpiperidine + NADPH + H+
-
725098
Withania coagulans
? + NADP+
-
-
-
?
1.1.1.206
4-ethylcyclohexanone + NADPH + H+
-
725098
Withania coagulans
4-ethylcyclohexanol + NADP+
-
-
-
?
1.1.1.206
4-methylcyclohexanone + NADPH + H+
-
725098
Withania coagulans
4-methylcyclohexanol + NADP+
-
-
-
?
1.1.1.206
4-piperidone + NADPH + H+
-
725098
Withania coagulans
? + NADP+
-
-
-
?
1.1.1.206
exo-6-hydroxytropinone + NADPH + H+
-
725098
Withania coagulans
? + NADP+
-
-
-
?
1.1.1.206
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
725098
Withania coagulans
?
-
-
-
?
1.1.1.206
additional information
no activity by TR-I with 1-methyl-2-pyrrolidone
725098
Withania coagulans
?
-
-
-
?
1.1.1.206
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
725098
Withania coagulans
?
-
-
-
?
1.1.1.206
N-methyl-4-piperidone + NADPH + H+
-
725098
Withania coagulans
? + NADP+
-
-
-
?
1.1.1.206
N-methylpiperidine + NADPH + H+
-
725098
Withania coagulans
? + NADP+
-
-
-
?
1.1.1.206
tetrahydro-4H-thiopyran-4-one + NADPH + H+
-
725098
Withania coagulans
? + NADP+
-
-
-
?
1.1.1.206
tropine + NADP+
in vitro reaction kinetics show that the enzyme predominantly favours the reverse reaction
725098
Withania coagulans
tropinone + NADPH + H+
-
-
-
r
1.1.1.206
tropinone + NADPH + H+
best substrate
725098
Withania coagulans
tropine + NADP+
-
-
-
r
1.1.1.206
tropinone + NADPH + H+
preferred substrate
725098
Withania coagulans
tropine + NADP+
-
-
-
r
1.1.1.206
tropinone + NADPH + H+
the reaction direction of tropinone reduction is predominantly operational in planta
725098
Withania coagulans
tropine + NADP+
-
-
-
r
1.1.1.206
tropinone + NADPH + H+
the in vitro reaction kinetics predominantly favour the reaction direction of tropinone reduction
725098
Withania coagulans
tropine + NADP+
-
-
-
r
Subunits (protein specific)
EC Number
Subunits
Commentary
Organism
1.1.1.206
?
x * 29340, sequence calculation
Withania coagulans
1.1.1.206
More
superimposition of three-dimensional models of tropinone reductases,overview
Withania coagulans
1.1.1.206
More
a three dimensional model is prepared by taking Datura stramonium TR-II, PDB ID 1ipf, as template, structure comparison, overview
Withania coagulans
Temperature Optimum [°C] (protein specific)
EC Number
Temperature Optimum [°C]
Temperature Optimum Maximum [°C]
Commentary
Organism
1.1.1.206
30
-
assay at
Withania coagulans
Temperature Stability [°C] (protein specific)
EC Number
Temperature Stability Minimum [°C]
Temperature Stability Maximum [°C]
Commentary
Organism
1.1.1.206
50
-
purified recombinant enzyme, 30 min, over 92% of maximal activity remaining
Withania coagulans
1.1.1.206
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
1.1.1.206
60
-
purified recombinant enzyme, 30 min, 30% of maximal activity remaining
Withania coagulans
1.1.1.206
70
-
purified recombinant enzyme, 30 min, 7% of maximal activity remaining
Withania coagulans
1.1.1.206
80
-
purified recombinant enzyme, 30 min, inactivation
Withania coagulans
Turnover Number [1/s] (protein specific)
EC Number
Turnover Number Minimum [1/s]
Turnover Number Maximum [1/s]
Substrate
Commentary
Organism
Structure
1.1.1.206
1.87
-
Tropine
pH 6.6, 30°C, recombinant enzyme
Withania coagulans
1.1.1.206
1.87
-
Tropine
pH 6.6, 30°C, recombinant His-tagged enzyme
Withania coagulans
1.1.1.206
2.94
-
NADP+
pH 6.6, 30°C, recombinant enzyme
Withania coagulans
1.1.1.206
2.94
-
NADP+
pH 6.6, 30°C, recombinant His-tagged enzyme
Withania coagulans
1.1.1.206
6.76
-
NADPH
pH 6.6, 30°C, recombinant enzyme
Withania coagulans
1.1.1.206
6.76
-
NADPH
pH 6.6, 30°C, recombinant His-tagged enzyme
Withania coagulans
1.1.1.206
16.74
-
tropinone
pH 6.6, 30°C, recombinant enzyme
Withania coagulans
1.1.1.206
16.74
-
tropinone
pH 6.6, 30°C, recombinant His-tagged enzyme
Withania coagulans
pH Optimum (protein specific)
EC Number
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
1.1.1.206
6.6
-
-
Withania coagulans
1.1.1.206
6.6
-
tropinone reduction
Withania coagulans
pH Range (protein specific)
EC Number
pH Minimum
pH Maximum
Commentary
Organism
1.1.1.206
additional information
-
narrow pH range in tropinone reduction
Withania coagulans
1.1.1.206
6.1
7
narrow pH range
Withania coagulans
pI Value (protein specific)
EC Number
Organism
Commentary
pI Value Maximum
pI Value
1.1.1.206
Withania coagulans
sequence calculation
-
6.12
General Information
EC Number
General Information
Commentary
Organism
1.1.1.206
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
1.1.1.206
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
1.1.1.206
metabolism
the enzyme catalyzes a step intermediary enroute to biosynthesis of tropane esters of medicinal importance, i.e. hyoscyamine/scopolamine
Withania coagulans
1.1.1.206
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
1.1.1.206
additional information
superimposition of three-dimensional models of tropinone reductases,overview
Withania coagulans
1.1.1.206
additional information
a three dimensional model is prepared by taking Datura stramonium TR-II, PDB ID 1ipf, as template, structure comparison, overview
Withania coagulans
General Information (protein specific)
EC Number
General Information
Commentary
Organism
1.1.1.206
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
1.1.1.206
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
1.1.1.206
metabolism
the enzyme catalyzes a step intermediary enroute to biosynthesis of tropane esters of medicinal importance, i.e. hyoscyamine/scopolamine
Withania coagulans
1.1.1.206
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
1.1.1.206
additional information
superimposition of three-dimensional models of tropinone reductases,overview
Withania coagulans
1.1.1.206
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 [mM/s]
EC Number
kcat/KM Value [1/mMs-1]
kcat/KM Value Maximum [1/mMs-1]
Substrate
Commentary
Organism
Structure
1.1.1.206
0.071
-
Tropine
pH 6.6, 30°C, recombinant enzyme
Withania coagulans
1.1.1.206
0.071
-
Tropine
pH 6.6, 30°C, recombinant His-tagged enzyme
Withania coagulans
1.1.1.206
11.53
-
tropinone
pH 6.6, 30°C, recombinant enzyme
Withania coagulans
1.1.1.206
11.53
-
tropinone
pH 6.6, 30°C, recombinant His-tagged enzyme
Withania coagulans
1.1.1.206
14
-
NADP+
pH 6.6, 30°C, recombinant enzyme
Withania coagulans
1.1.1.206
14
-
NADP+
pH 6.6, 30°C, recombinant His-tagged enzyme
Withania coagulans
1.1.1.206
53
-
NADPH
pH 6.6, 30°C, recombinant enzyme
Withania coagulans
1.1.1.206
53
-
NADPH
pH 6.6, 30°C, recombinant His-tagged enzyme
Withania coagulans
KCat/KM [mM/s] (protein specific)
EC Number
KCat/KM Value [1/mMs-1]
KCat/KM Value Maximum [1/mMs-1]
Substrate
Commentary
Organism
Structure
1.1.1.206
0.071
-
Tropine
pH 6.6, 30°C, recombinant enzyme
Withania coagulans
1.1.1.206
0.071
-
Tropine
pH 6.6, 30°C, recombinant His-tagged enzyme
Withania coagulans
1.1.1.206
11.53
-
tropinone
pH 6.6, 30°C, recombinant enzyme
Withania coagulans
1.1.1.206
11.53
-
tropinone
pH 6.6, 30°C, recombinant His-tagged enzyme
Withania coagulans
1.1.1.206
14
-
NADP+
pH 6.6, 30°C, recombinant enzyme
Withania coagulans
1.1.1.206
14
-
NADP+
pH 6.6, 30°C, recombinant His-tagged enzyme
Withania coagulans
1.1.1.206
53
-
NADPH
pH 6.6, 30°C, recombinant enzyme
Withania coagulans
1.1.1.206
53
-
NADPH
pH 6.6, 30°C, recombinant His-tagged enzyme
Withania coagulans