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Literature summary for 1.5.3.5 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)

Crystallization (Comment) Organism
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

Protein Variants Comment Organism
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
N462Y/W427Y mutant enzyme shows increased catalytic activity Pseudomonas putida
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]

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

Organism

Organism UniProt Comment Textmining
Pseudomonas putida F8G0P2
-
-
Pseudomonas putida S16 F8G0P2
-
-

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
(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
-
?
(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

Synonyms Comment Organism
flavoprotein nicotine oxidoreductase
-
Pseudomonas putida
NicA2
-
Pseudomonas putida

Cofactor

Cofactor Comment Organism Structure
FAD conserved FAD-binding domain Pseudomonas putida