BRENDA - Enzyme Database

Biochemical and molecular characterizations of nicotine demethylase in tobacco

Xu, D.; Shen, Y.; Chappell, J.; Cui, M.; Nielsen, M.T.; Physiol. Plant. 129, 307-319 (2007)
No PubMed abstract available

Data extracted from this reference:

Cloned(Commentary)
EC Number
Commentary
Organism
1.14.14.B11
D121-AA8 cDNA transformed into yeast cells; expression in yeast
Nicotiana tabacum
Inhibitors
EC Number
Inhibitors
Commentary
Organism
Structure
1.14.14.B11
1-aminobenzotriazole
at 1 mM inhibits by 17%
Nicotiana tabacum
1.14.14.B11
Chloropromazine
1 mM, 87% inhibition of recombinant nicotine demethylase activity in yeast microsomes
Nicotiana tabacum
1.14.14.B11
chlorpromazine
at 1 mM inhibits by 87%
Nicotiana tabacum
1.14.14.B11
CO
partial reverse activity after light exposure
Nicotiana tabacum
1.14.14.B11
cytochrome c
1 mM, 78% inhibition of recombinant nicotine demethylase activity in yeast microsomes
Nicotiana tabacum
1.14.14.B11
oxidized cytochrome c
at 0.1 mM inhibits by 78%
Nicotiana tabacum
1.14.14.B11
Piperonylbutoxide
at 1 mM inhibits by 42%
Nicotiana tabacum
KM Value [mM]
EC Number
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
1.14.14.B11
0.0039
-
Nicotine
; at 25°C, pH 6.25
Nicotiana tabacum
Localization
EC Number
Localization
Commentary
Organism
GeneOntology No.
Textmining
1.14.14.B11
membrane
putative N-terminal hydrophobic trans-membrane domain
Nicotiana tabacum
16020
-
1.14.14.B11
microsome
-
Nicotiana tabacum
-
-
Metals/Ions
EC Number
Metals/Ions
Commentary
Organism
Structure
1.14.14.B11
Iron
cysteine-containing heme-binding region
Nicotiana tabacum
Natural Substrates/ Products (Substrates)
EC Number
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
1.14.14.B11
nicotine + NADPH + O2 + H+
Nicotiana tabacum
demethylates nicotine to form nornicotine, a precursor to the nitrosamine N'-nitrosonornicotine
nornicotine + formaldehyde + NADP+ + H2O
-
-
?
Organism
EC Number
Organism
Primary Accession No. (UniProt)
Commentary
Textmining
1.14.14.B11
Nicotiana tabacum
Q38Q87
; var. 4407
-
Source Tissue
EC Number
Source Tissue
Commentary
Organism
Textmining
1.14.14.B11
leaf
; induced in leaves treated with the growth regulator ethylene
Nicotiana tabacum
-
Substrates and Products (Substrate)
EC Number
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
1.14.14.B11
nicotine + NADPH + O2 + H+
-
700634
Nicotiana tabacum
nornicotine + formaldehyde + NADP+ + H2O
-
-
-
?
1.14.14.B11
nicotine + NADPH + O2 + H+
demethylates nicotine to form nornicotine, a precursor to the nitrosamine N'-nitrosonornicotine
700634
Nicotiana tabacum
nornicotine + formaldehyde + NADP+ + H2O
-
-
-
?
1.14.14.B11
nicotine + NADPH + O2 + H+
six putative substrate recognition sites
700634
Nicotiana tabacum
nornicotine + formaldehyde + NADP+ + H2O
-
-
-
?
Cofactor
EC Number
Cofactor
Commentary
Organism
Structure
1.14.14.B11
cytochrome P450
-
Nicotiana tabacum
1.14.14.B11
NADPH
; is required for full activity
Nicotiana tabacum
Cloned(Commentary) (protein specific)
EC Number
Commentary
Organism
1.14.14.B11
D121-AA8 cDNA transformed into yeast cells; expression in yeast
Nicotiana tabacum
Cofactor (protein specific)
EC Number
Cofactor
Commentary
Organism
Structure
1.14.14.B11
cytochrome P450
-
Nicotiana tabacum
1.14.14.B11
NADPH
; is required for full activity
Nicotiana tabacum
Inhibitors (protein specific)
EC Number
Inhibitors
Commentary
Organism
Structure
1.14.14.B11
1-aminobenzotriazole
at 1 mM inhibits by 17%
Nicotiana tabacum
1.14.14.B11
Chloropromazine
1 mM, 87% inhibition of recombinant nicotine demethylase activity in yeast microsomes
Nicotiana tabacum
1.14.14.B11
chlorpromazine
at 1 mM inhibits by 87%
Nicotiana tabacum
1.14.14.B11
CO
partial reverse activity after light exposure
Nicotiana tabacum
1.14.14.B11
cytochrome c
1 mM, 78% inhibition of recombinant nicotine demethylase activity in yeast microsomes
Nicotiana tabacum
1.14.14.B11
oxidized cytochrome c
at 0.1 mM inhibits by 78%
Nicotiana tabacum
1.14.14.B11
Piperonylbutoxide
at 1 mM inhibits by 42%
Nicotiana tabacum
KM Value [mM] (protein specific)
EC Number
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
1.14.14.B11
0.0039
-
Nicotine
; at 25°C, pH 6.25
Nicotiana tabacum
Localization (protein specific)
EC Number
Localization
Commentary
Organism
GeneOntology No.
Textmining
1.14.14.B11
membrane
putative N-terminal hydrophobic trans-membrane domain
Nicotiana tabacum
16020
-
1.14.14.B11
microsome
-
Nicotiana tabacum
-
-
Metals/Ions (protein specific)
EC Number
Metals/Ions
Commentary
Organism
Structure
1.14.14.B11
Iron
cysteine-containing heme-binding region
Nicotiana tabacum
Natural Substrates/ Products (Substrates) (protein specific)
EC Number
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
1.14.14.B11
nicotine + NADPH + O2 + H+
Nicotiana tabacum
demethylates nicotine to form nornicotine, a precursor to the nitrosamine N'-nitrosonornicotine
nornicotine + formaldehyde + NADP+ + H2O
-
-
?
Source Tissue (protein specific)
EC Number
Source Tissue
Commentary
Organism
Textmining
1.14.14.B11
leaf
; induced in leaves treated with the growth regulator ethylene
Nicotiana tabacum
-
Substrates and Products (Substrate) (protein specific)
EC Number
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
1.14.14.B11
nicotine + NADPH + O2 + H+
-
700634
Nicotiana tabacum
nornicotine + formaldehyde + NADP+ + H2O
-
-
-
?
1.14.14.B11
nicotine + NADPH + O2 + H+
demethylates nicotine to form nornicotine, a precursor to the nitrosamine N'-nitrosonornicotine
700634
Nicotiana tabacum
nornicotine + formaldehyde + NADP+ + H2O
-
-
-
?
1.14.14.B11
nicotine + NADPH + O2 + H+
six putative substrate recognition sites
700634
Nicotiana tabacum
nornicotine + formaldehyde + NADP+ + H2O
-
-
-
?
Expression
EC Number
Organism
Commentary
Expression
1.14.14.B11
Nicotiana tabacum
ethylene induces microsomal demethylase activity in leaves
up
General Information
EC Number
General Information
Commentary
Organism
1.14.14.B11
physiological function
only microsomes from yeast expressing the D121-AA8 cDNA exhibit significant nicotine demethylase activity
Nicotiana tabacum
General Information (protein specific)
EC Number
General Information
Commentary
Organism
1.14.14.B11
physiological function
only microsomes from yeast expressing the D121-AA8 cDNA exhibit significant nicotine demethylase activity
Nicotiana tabacum
Expression (protein specific)
EC Number
Organism
Commentary
Expression
1.14.14.B11
Nicotiana tabacum
ethylene induces microsomal demethylase activity in leaves
up