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Literature summary extracted from

  • Tararina, M.A.; Dam, K.K.; Dhingra, M.; Janda, K.D.; Palfey, B.A.; Allen, K.N.
    Fast kinetics reveals rate-limiting oxidation and the role of the aromatic cage in the mechanism of the nicotine-degrading enzyme NicA2 (2021), Biochemistry, 60, 259-273 .
    View publication on PubMed

Crystallization (Commentary)

EC Number Crystallization (Comment) Organism
1.5.3.5 hanging-drop vapor diffusion method, X-ray crystal structures of the N462V and N462Y/W427Y variants complexed with (S)-nicotine (at 2.7 and 2.3 A resolution, respectively) reveal no significant active-site rearrangements Pseudomonas putida

Protein Variants

EC Number Protein Variants Comment Organism
1.5.3.5 N462H the variant shows moderately higher oxidase activity, reductive half-reaction using (S)-nicotine is significantly slower than that of wild-type enzyme Pseudomonas putida
1.5.3.5 N462Y/W427Y mutant enzyme shows increased catalytic activity Pseudomonas putida
1.5.3.5 W427V the variant shows moderately higher oxidase activity, reductive half-reaction using (S)-nicotine is significantly slower than that of wild-type enzyme Pseudomonas putida

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
1.5.3.5 0.000114
-
 (S)-nicotine pH and temperature not specified in the publication Pseudomonas putida

Organism

EC Number Organism UniProt Comment Textmining
1.5.3.5 Pseudomonas putida F8G0P2
-
-
1.5.3.5 Pseudomonas putida S16 F8G0P2
-
-

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
1.5.3.5 (S)-nicotine + H2O + O2 N-methyl-myosmine, which is nonenzymatically hydrolyzed to pseudooxynicotine. The flavoenzyme preferentially oxidizes tertiary amines with an efficient reductive half-reaction and a very slow oxidative half-reaction when O2 is the oxidizing substrate, suggesting that the true oxidizing agent is unknown Pseudomonas putida N-methylmyosmine + H2O2
-
 ?
1.5.3.5 (S)-nicotine + H2O + O2 N-methyl-myosmine, which is nonenzymatically hydrolyzed to pseudooxynicotine. The flavoenzyme preferentially oxidizes tertiary amines with an efficient reductive half-reaction and a very slow oxidative half-reaction when O2 is the oxidizing substrate, suggesting that the true oxidizing agent is unknown Pseudomonas putida S16 N-methylmyosmine + H2O2
-
 ?

Synonyms

EC Number Synonyms Comment Organism
1.5.3.5 flavoprotein nicotine oxidoreductase
-
 Pseudomonas putida
1.5.3.5 NicA2
-
 Pseudomonas putida

Cofactor

EC Number Cofactor Comment Organism Structure
1.5.3.5 FAD conserved FAD-binding domain Pseudomonas putida