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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
Search for more literature for 1.5.3.5

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
a

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