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Literature summary for 1.5.3.5 extracted from

  • Fitzpatrick, P.F.; Chadegani, F.; Zhang, S.; Dougherty, V.
    Mechanism of flavoprotein L-6-hydroxynicotine oxidase pH and solvent isotope effects and identification of key active site residues (2017), Biochemistry, 56, 869-875 .
    View publication on PubMedView publication on EuropePMC

Cloned(Commentary)

Cloned (Comment) Organism
expression in Escherichi coli Paenarthrobacter nicotinovorans

Protein Variants

Protein Variants Comment Organism
K287M mutation results in an about 10-fold decreases in kcat/Km and k(red) for (S)-6-hydroxynicotine and a 6000-fold decrease in the kcat/Km value for oxygen Paenarthrobacter nicotinovorans
N166A mutation results in an about 30fold decrease in kcat/Km and k(red) for (S)-6-hydroxynicotine, respectively, with larger effects on the kcat/Km value for (S)-6-hydroxynornicotine. The shapes of the pH profiles are not altered Paenarthrobacter nicotinovorans
Y311F mutation results in an about 30fold decrease in kcat/Km and k(red) for (S)-6-hydroxynicotine, respectively, with larger effects on the kcat/Km value for (S)-6-hydroxynornicotine. The shapes of the pH profiles are not altered Paenarthrobacter nicotinovorans

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.011
-
(S)-6-hydroxynicotine pH 7.0, 25°C, mutant enzyme K287M Paenarthrobacter nicotinovorans
0.064
-
(S)-6-hydroxynornicotine pH 7.0, 25°C, wild-type enzyme Paenarthrobacter nicotinovorans
0.13
-
(S)-6-hydroxynicotine pH 7.0, 25°C, wild-type enzyme Paenarthrobacter nicotinovorans
0.19
-
O2 pH 7.0, 25°C, mutant enzyme Y311F, cosubstrate: (S)-6-hydroxynicotine Paenarthrobacter nicotinovorans
0.2
-
O2 pH 7.0, 25°C, mutant enzyme N166A, cosubstrate: (S)-6-hydroxynicotine Paenarthrobacter nicotinovorans
0.2
-
(S)-6-hydroxynicotine pH 7.0, 25°C, mutant enzyme Y311F Paenarthrobacter nicotinovorans
0.29
-
O2 pH 7.0, 25°C, wild-type enzyme, cosubstrate: (S)-6-hydroxynicotine Paenarthrobacter nicotinovorans
0.34
-
(S)-6-hydroxynicotine pH 7.0, 25°C, mutant enzyme N166A Paenarthrobacter nicotinovorans
0.46
-
O2 pH 7.0, 25°C, wild-type enzyme, cosubstrate: (S)-6-hydroxynornicotine Paenarthrobacter nicotinovorans
1.1
-
(S)-6-hydroxynornicotine pH 7.0, 25°C, mutant enzyme Y311F Paenarthrobacter nicotinovorans
1.2
-
(S)-6-hydroxynornicotine pH 7.0, 25°C, mutant enzyme N166A Paenarthrobacter nicotinovorans

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
(S)-6-hydroxynicotine + H2O + O2 Paenarthrobacter nicotinovorans the enzyme catalyzes the oxidation of (S)-6-hydroxynicotine to 6-hydroxypseudooxynicotine during microbial catabolism of nicotine 1-(6-hydroxypyridin-3-yl)-4-(methylamino)butan-1-one + H2O2
-
?

Organism

Organism UniProt Comment Textmining
Paenarthrobacter nicotinovorans
-
-
-

Purification (Commentary)

Purification (Comment) Organism
-
Paenarthrobacter nicotinovorans

Reaction

Reaction Comment Organism Reaction ID
(S)-6-hydroxynicotine + H2O + O2 = 1-(6-hydroxypyridin-3-yl)-4-(methylamino)butan-1-one + H2O2 mechanism and roles of active site residues: Asn166 and Tyr311 bind the substrate, while Lys287 forms a water-mediated hydrogen bond with flavin N5 Paenarthrobacter nicotinovorans

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
(S)-6-hydroxynicotine + H2O + O2
-
Paenarthrobacter nicotinovorans 1-(6-hydroxypyridin-3-yl)-4-(methylamino)butan-1-one + H2O2
-
?
(S)-6-hydroxynicotine + H2O + O2 the enzyme catalyzes the oxidation of (S)-6-hydroxynicotine to 6-hydroxypseudooxynicotine during microbial catabolism of nicotine Paenarthrobacter nicotinovorans 1-(6-hydroxypyridin-3-yl)-4-(methylamino)butan-1-one + H2O2
-
?
(S)-6-hydroxynornicotine + H2O + O2
-
Paenarthrobacter nicotinovorans ?
-
?

Synonyms

Synonyms Comment Organism
L-6-hydroxynicotine oxidase
-
Paenarthrobacter nicotinovorans
LHNO
-
Paenarthrobacter nicotinovorans

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
25
-
assay at Paenarthrobacter nicotinovorans

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
0.26
-
(S)-6-hydroxynicotine pH 7.0, 25°C, mutant enzyme K287M Paenarthrobacter nicotinovorans
2.5
-
(S)-6-hydroxynornicotine pH 7.0, 25°C, mutant enzyme N166A Paenarthrobacter nicotinovorans
4.8
-
(S)-6-hydroxynicotine pH 7.0, 25°C, mutant enzyme N166A Paenarthrobacter nicotinovorans
9.1
-
(S)-6-hydroxynornicotine pH 7.0, 25°C, mutant enzyme Y311F Paenarthrobacter nicotinovorans
16
-
(S)-6-hydroxynornicotine pH 7.0, 25°C, wild-type enzyme Paenarthrobacter nicotinovorans
20.9
-
(S)-6-hydroxynicotine pH 7.0, 25°C, mutant enzyme Y311F Paenarthrobacter nicotinovorans
78
-
(S)-6-hydroxynicotine pH 7.0, 25°C, wild-type enzyme Paenarthrobacter nicotinovorans

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
8
-
assay at Paenarthrobacter nicotinovorans

Cofactor

Cofactor Comment Organism Structure
FAD flavoprotein Paenarthrobacter nicotinovorans

General Information

General Information Comment Organism
metabolism the enzyme catalyzes the oxidation of (S)-6-hydroxynicotine to 6-hydroxypseudooxynicotine during microbial catabolism of nicotine Paenarthrobacter nicotinovorans

kcat/KM [mM/s]

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
0.048
-
O2 pH 7.0, 25°C, mutant enzyme K287M, cosubstrate: (S)-6-hydroxynicotine Paenarthrobacter nicotinovorans
2
-
(S)-6-hydroxynornicotine pH 7.0, 25°C, mutant enzyme N166A Paenarthrobacter nicotinovorans
8.6
-
(S)-6-hydroxynornicotine pH 7.0, 25°C, mutant enzyme Y311F Paenarthrobacter nicotinovorans
14
-
(S)-6-hydroxynicotine pH 7.0, 25°C, mutant enzyme N166A Paenarthrobacter nicotinovorans
24
-
(S)-6-hydroxynicotine pH 7.0, 25°C, mutant enzyme K287M Paenarthrobacter nicotinovorans
24
-
O2 pH 7.0, 25°C, mutant enzyme N166A, cosubstrate: (S)-6-hydroxynicotine Paenarthrobacter nicotinovorans
103
-
(S)-6-hydroxynicotine pH 7.0, 25°C, mutant enzyme Y311F Paenarthrobacter nicotinovorans
110
-
O2 pH 7.0, 25°C, mutant enzyme Y311F, cosubstrate: (S)-6-hydroxynicotine Paenarthrobacter nicotinovorans
150
-
O2 pH 7.0, 25°C, wild-type enzyme, cosubstrate: (S)-6-hydroxynornicotine Paenarthrobacter nicotinovorans
270
-
O2 pH 7.0, 25°C, wild-type enzyme, cosubstrate: (S)-6-hydroxynicotine Paenarthrobacter nicotinovorans
370
-
(S)-6-hydroxynornicotine pH 7.0, 25°C, wild-type enzyme Paenarthrobacter nicotinovorans
600
-
(S)-6-hydroxynicotine pH 7.0, 25°C, wild-type enzyme Paenarthrobacter nicotinovorans