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Literature summary extracted from

  • Kushwaha, A.K.; Sangwan, N.S.; Trivedi, P.K.; Negi, A.S.; Misra, L.; Sangwan, R.S.
    Tropine forming tropinone reductase gene from Withania somnifera (Ashwagandha): biochemical characteristics of the recombinant enzyme and novel physiological overtones of tissue-wide gene expression patterns (2013), PLoS ONE, 8, e74777.
    View publication on PubMedView publication on EuropePMC

Cloned(Commentary)

EC Number Cloned (Comment) Organism
1.1.1.206 gene TR-I from roots, DNA and amino acid sequence determination and analysis, sequence comparisons and phylogenetic analysis and tree, recombinant functional overexpression of His6-tagged enzyme in Escherichia coli strain BL21(DE3) Withania somnifera

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
1.1.1.206 additional information
-
 additional information Michaelis-Menten kinetics Withania somnifera
1.1.1.206 0.00043
-
 NADPH recombinant enzyme, pH 6.7, 30°C Withania somnifera
1.1.1.206 0.0962
-
 NADP+ recombinant enzyme, pH 6.7, 30°C Withania somnifera
1.1.1.206 1.27
-
 tropinone recombinant enzyme, pH 6.7, 30°C Withania somnifera
1.1.1.206 1.74
-
 N-methylpiperidine recombinant enzyme, pH 6.7, 30°C Withania somnifera
1.1.1.206 2.11
-
 3-methylcyclohexanone recombinant enzyme, pH 6.7, 30°C Withania somnifera
1.1.1.206 114.9
-
 Tropine recombinant enzyme, pH 6.7, 30°C Withania somnifera

Metals/Ions

EC Number Metals/Ions Comment Organism Structure
1.1.1.206 Mg2+ slightly activating Withania somnifera
1.1.1.206 additional information except slight enhancement of enzyme activity by Mg2+ and Na+, none of the following cations, Mn2+, Li+, NH4+, Mg2+, Na+, and Co2+, 5 mM each, affects the activity significantly Withania somnifera
1.1.1.206 Na+ slightly activating Withania somnifera

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
1.1.1.206 60000
-
 2 * 60000, SDS-PAGE Withania somnifera

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+ Withania somnifera
-
 tropine + NADP+
-
 r

Organism

EC Number Organism UniProt Comment Textmining
1.1.1.206 Withania somnifera U5PUZ1 cv. NMITLI-118
-

Purification (Commentary)

EC Number Purification (Comment) Organism
1.1.1.206 recombinant His6-tagged enzyme from Escherichia coli strain BL21(DE3) by affinity chromatography and gel filtration Withania somnifera

Source Tissue

EC Number Source Tissue Comment Organism Textmining
1.1.1.206 berry low enzyme level, all developmental stages Withania somnifera
-
1.1.1.206 leaf low enzyme level, all developmental stages Withania somnifera
-
1.1.1.206 additional information enzyme activity and tropine/tropinone contents during different developmental stages, overview Withania somnifera
-
1.1.1.206 root high enzyme level in bark, moderate in medulla Withania somnifera
-
1.1.1.206 shoot high enzyme level Withania somnifera
-
1.1.1.206 stem high enzyme level in bark, moderate in medulla Withania somnifera
-

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
1.1.1.206 1-propyl-4-piperidone + NADPH + H+ low activity Withania somnifera 1-propylpiperidin-4-ol + NADP+
-
 ?
1.1.1.206 2-pyrrolidone + NADPH + H+ very low activity Withania somnifera 2-hydroxypyrrolidine + NADP+
-
 ?
1.1.1.206 3-methylcyclohexanone + NADPH + H+
-
 Withania somnifera 3-methylcyclohexanol + NADP+
-
 ?
1.1.1.206 4-chloro-1-methylpiperidine + NADPH + H+ moderate activity Withania somnifera ? + NADP+
-
 ?
1.1.1.206 4-ethylcyclohexanone + NADPH + H+ low activity Withania somnifera 4-ethylcyclohexanol + NADP+
-
 ?
1.1.1.206 4-methylcyclohexanone + NADPH + H+ low activity Withania somnifera 4-methylcyclohexanol + NADP+
-
 ?
1.1.1.206 4-piperidone + NADPH + H+ very low activity Withania somnifera 4-hydroxypiperidin + NADP+
-
 ?
1.1.1.206 exo-6-hydroxytropinone + NADPH + H+ low activity Withania somnifera ? + NADP+
-
 ?
1.1.1.206 additional information TLC and GC-MS analysis of substrates and products. The enzyme also shows comparatively efficient catalysis of NADPH-dependent reduction of cyclic ketones 3-methylcyclohexanone and N-methylpiperidine, substrate specificity, overview. No activity with 1-methyl-2-pyrrolidone Withania somnifera ?
-
 ?
1.1.1.206 N-methyl-4-piperidone + NADPH + H+ low activity Withania somnifera 1-methylpiperidin-4-ol + NADP+
-
 ?
1.1.1.206 N-methylpiperidine + NADPH + H+
-
 Withania somnifera ? + NADP+
-
 ?
1.1.1.206 tetrahydro-4H-thiopyran-4-one + NADPH + H+ moderate activity Withania somnifera ? + NADP+
-
 ?
1.1.1.206 tropine + NADP+ catalytic kinetics of the enzyme favour the forward reaction, tropine formation Withania somnifera tropinone + NADPH + H+
-
 r
1.1.1.206 tropinone + NADPH + H+
-
 Withania somnifera tropine + NADP+
-
 r

Subunits

EC Number Subunits Comment Organism
1.1.1.206 homodimer 2 * 60000, SDS-PAGE Withania somnifera
1.1.1.206 More the homodimer is the catalytically active form of the recombinant WsTR-I Withania somnifera

Synonyms

EC Number Synonyms Comment Organism
1.1.1.206 TR-I
-
 Withania somnifera
1.1.1.206 tropinone reductase
-
 Withania somnifera
1.1.1.206 tropinone reductase-I
-
 Withania somnifera
1.1.1.206 WsTR-I
-
 Withania somnifera
1.1.1.206 WsTR-I protein
-
 Withania somnifera

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
1.1.1.206 30
-
 assay at Withania somnifera

Temperature Stability [°C]

EC Number Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
1.1.1.206 50 60 purified recombinant enzyme, retains about 75% of maximal catalytic activity at up to 50°C, 4.27% activity remaining on incubation at 60°C Withania somnifera

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
1.1.1.206 0.54
-
 NADP+ recombinant enzyme, pH 6.7, 30°C Withania somnifera
1.1.1.206 1.67
-
 Tropine recombinant enzyme, pH 6.7, 30°C Withania somnifera
1.1.1.206 8.97
-
 3-methylcyclohexanone recombinant enzyme, pH 6.7, 30°C Withania somnifera
1.1.1.206 9.23
-
 N-methylpiperidine recombinant enzyme, pH 6.7, 30°C Withania somnifera
1.1.1.206 9.51
-
 tropinone recombinant enzyme, pH 6.7, 30°C Withania somnifera
1.1.1.206 33.34
-
 NADPH recombinant enzyme, pH 6.7, 30°C Withania somnifera

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
1.1.1.206 6.7
-
-
 Withania somnifera

Cofactor

EC Number Cofactor Comment Organism Structure
1.1.1.206 NADP+
-
 Withania somnifera
1.1.1.206 NADPH
-
 Withania somnifera

Expression

EC Number Organism Comment Expression
1.1.1.206 Withania somnifera the enzyme-encoding TR-I gene expression is upregulated on exposure to signal molecules, i.e. methyl jasmonate and salicylic acid, and on mechanical injury/wounding up

General Information

EC Number General Information Comment Organism
1.1.1.206 evolution molecular evolution of WsTR-I, overview Withania somnifera
1.1.1.206 metabolism the enzyme catalyzes the NADPH-dependent tropinone to tropine conversion step in tropane metabolism, metabolic pathway of tropane alkaloid biosynthesis, overview Withania somnifera
1.1.1.206 additional information enzyme three-dimensional structure modelling Withania somnifera
1.1.1.206 physiological function physiological role of tropinone reductase enzyme in tropane alkaloid biosynthesis in aerial tissues of the plant. The metabolic step of tropine formation may be regulated by, besides through the transcript and protein levels, the tissue concentration of tropinone Withania somnifera

kcat/KM [mM/s]

EC Number kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
1.1.1.206 0.014
-
 Tropine recombinant enzyme, pH 6.7, 30°C Withania somnifera
1.1.1.206 0.559
-
 NADP+ recombinant enzyme, pH 6.7, 30°C Withania somnifera
1.1.1.206 4.38
-
 N-methylpiperidine recombinant enzyme, pH 6.7, 30°C Withania somnifera
1.1.1.206 5.17
-
 3-methylcyclohexanone recombinant enzyme, pH 6.7, 30°C Withania somnifera
1.1.1.206 7.49
-
 tropinone recombinant enzyme, pH 6.7, 30°C Withania somnifera
1.1.1.206 77540
-
 NADPH recombinant enzyme, pH 6.7, 30°C Withania somnifera