Protein Variants | Comment | Organism |
---|---|---|
E350A | substitution decreases the kcat/Kamine value by about 20fold | Paenarthrobacter nicotinovorans |
E350Q | substitution decreases the kcat/Kamine value by about 220fold | Paenarthrobacter nicotinovorans |
E352Q | substitution decreases the kcat/Kamine value about 3800fold | Paenarthrobacter nicotinovorans |
K348M | substitution has only a small effect on the kinetic parameters | Paenarthrobacter nicotinovorans |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.2 | - |
(R)-6-hydroxynicotine | wild-type, pH 8, 25°C | Paenarthrobacter nicotinovorans | |
0.2 | - |
O2 | mutant E352Q, pH 8, 25°C | Paenarthrobacter nicotinovorans | |
0.21 | - |
(R)-6-chloronicotine | wild-type, pH 8, 25°C | Paenarthrobacter nicotinovorans | |
0.32 | - |
(R)-6-hydroxynicotine | mutant K348M, pH 8, 25°C | Paenarthrobacter nicotinovorans | |
0.34 | - |
(R)-6-hydroxynornicotine | wild-type, pH 8, 25°C | Paenarthrobacter nicotinovorans | |
0.36 | - |
(R)-4-(1-methylpyrrolidine-2-yl)phenol | wild-type, pH 8, 25°C | Paenarthrobacter nicotinovorans | |
0.43 | - |
O2 | mutant E350Q, pH 8, 25°C | Paenarthrobacter nicotinovorans | |
0.83 | - |
(R)-6-hydroxynornicotine | mutant K348M, pH 8, 25°C | Paenarthrobacter nicotinovorans | |
0.84 | - |
(R)-nicotine | wild-type, pH 8, 25°C | Paenarthrobacter nicotinovorans | |
1.12 | - |
(R)-6-hydroxynicotine | mutant E350A, pH 8, 25°C | Paenarthrobacter nicotinovorans | |
1.2 | - |
O2 | mutant K348M, pH 8, 25°C | Paenarthrobacter nicotinovorans | |
1.8 | - |
O2 | mutant E350A, pH 8, 25°C | Paenarthrobacter nicotinovorans | |
4.7 | - |
O2 | wild-type, pH 8, 25°C | Paenarthrobacter nicotinovorans | |
5.5 | - |
(R)-6-hydroxynicotine | mutant E350Q, pH 8, 25°C | Paenarthrobacter nicotinovorans | |
6 | - |
(R)-6-hydroxynicotine | mutant E352Q, pH 8, 25°C | Paenarthrobacter nicotinovorans |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Paenarthrobacter nicotinovorans | Q8GAG1 | - |
- |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
(R)-4-(1-methylpyrrolidine-2-yl)phenol + H2O + O2 | - |
Paenarthrobacter nicotinovorans | ? + H2O2 | - |
? | |
(R)-6-chloronicotine + H2O + O2 | - |
Paenarthrobacter nicotinovorans | 4-amino-1-(6-chloropyridin-3-yl)-butan-1-one + H2O2 | - |
? | |
(R)-6-hydroxynicotine + H2O + O2 | - |
Paenarthrobacter nicotinovorans | 1-(6-hydroxypyrid-3-yl)-4-(methylamino)-butan-1-one + H2O2 | - |
? | |
(R)-6-hydroxynornicotine + H2O + O2 | - |
Paenarthrobacter nicotinovorans | 4-amino-1-(6-hydroxypyridin-3-yl)-butan-1-one + H2O2 | - |
? | |
(R)-nicotine + H2O + O2 | - |
Paenarthrobacter nicotinovorans | 1-(pyrid-3-yl)-4-(methylamino)-butan-1-one + H2O2 | - |
? |
Subunits | Comment | Organism |
---|---|---|
? | x * 49000, calculated from sequence | Paenarthrobacter nicotinovorans |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.26 | - |
(R)-nicotine | mutant E350Q, pH 8, 25°C | Paenarthrobacter nicotinovorans | |
0.63 | - |
(R)-6-hydroxynicotine | mutant E352Q, pH 8, 25°C | Paenarthrobacter nicotinovorans | |
0.85 | - |
(R)-6-chloronicotine | mutant E350A, pH 8, 25°C | Paenarthrobacter nicotinovorans | |
1.5 | - |
(R)-4-(1-methylpyrrolidine-2-yl)phenol | mutant E352Q, pH 8, 25°C | Paenarthrobacter nicotinovorans | |
5.4 | - |
(R)-6-hydroxynornicotine | mutant K348M, pH 8, 25°C | Paenarthrobacter nicotinovorans | |
8.1 | - |
(R)-6-hydroxynornicotine | wild-type, pH 8, 25°C | Paenarthrobacter nicotinovorans | |
9.3 | - |
(R)-6-hydroxynicotine | mutant E350Q, pH 8, 25°C | Paenarthrobacter nicotinovorans | |
20 | - |
(R)-6-hydroxynicotine | mutant E350A, pH 8, 25°C | Paenarthrobacter nicotinovorans | |
46 | - |
(R)-6-hydroxynicotine | mutant K348M, pH 8, 25°C | Paenarthrobacter nicotinovorans | |
77 | - |
(R)-6-hydroxynicotine | wild-type, pH 8, 25°C | Paenarthrobacter nicotinovorans |
General Information | Comment | Organism |
---|---|---|
metabolism | a network of hydrogen-bonded residues in the active site is involved in binding the neutral form of the amine substrate, followed by the transfer of a hydride from the amine to the flavin. There is no solvent isotope effect on the kcat/Kamine value with either (R)-6-hydroxynicotine or the slower substrate (R)-6-hydroxynornicotine. Only the neutral form of the substrate and the correctly protonated form of the enzyme bind. The active-site residues Lys348, Glu350, and Glu352 are all properly positioned for substrate binding | Paenarthrobacter nicotinovorans |
kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.1 | - |
(R)-6-hydroxynicotine | mutant E352Q, pH 8, 25°C | Paenarthrobacter nicotinovorans | |
0.15 | - |
(R)-nicotine | wild-type, pH 8, 25°C | Paenarthrobacter nicotinovorans | |
1.7 | - |
(R)-6-hydroxynicotine | mutant E350Q, pH 8, 25°C | Paenarthrobacter nicotinovorans | |
3.2 | - |
O2 | mutant E352Q, pH 8, 25°C | Paenarthrobacter nicotinovorans | |
4 | - |
(R)-6-chloronicotine | wild-type, pH 8, 25°C | Paenarthrobacter nicotinovorans | |
4 | - |
(R)-4-(1-methylpyrrolidine-2-yl)phenol | wild-type, pH 8, 25°C | Paenarthrobacter nicotinovorans | |
6.5 | - |
(R)-6-hydroxynornicotine | mutant K348M, pH 8, 25°C | Paenarthrobacter nicotinovorans | |
11.5 | - |
O2 | mutant E350A, pH 8, 25°C | Paenarthrobacter nicotinovorans | |
16 | - |
O2 | wild-type, pH 8, 25°C | Paenarthrobacter nicotinovorans | |
17.8 | - |
(R)-6-hydroxynicotine | mutant E350A, pH 8, 25°C | Paenarthrobacter nicotinovorans | |
21 | - |
O2 | mutant E350Q, pH 8, 25°C | Paenarthrobacter nicotinovorans | |
24 | - |
(R)-6-hydroxynornicotine | wild-type, pH 8, 25°C | Paenarthrobacter nicotinovorans | |
39 | - |
O2 | mutant K348M, pH 8, 25°C | Paenarthrobacter nicotinovorans | |
144 | - |
(R)-6-hydroxynicotine | mutant K348M, pH 8, 25°C | Paenarthrobacter nicotinovorans | |
380 | - |
(R)-6-hydroxynicotine | wild-type, pH 8, 25°C | Paenarthrobacter nicotinovorans |