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Literature summary for 1.5.99.4 extracted from

  • Qiu, J.; Ma, Y.; Zhang, J.; Wen, Y.; Liu, W.
    Cloning of a novel nicotine oxidase gene from Pseudomonas sp. strain HZN6 whose product nonenantioselectively degrades nicotine to pseudooxynicotine (2013), Appl. Environ. Microbiol., 79, 2164-2171.
    View publication on PubMedView publication on EuropePMC

Cloned(Commentary)

Cloned (Comment) Organism
gene nox, DNA and amino acid sequence deteremination and analysis, recombinant expression from a broad-host-range cloning vector in Escherichia coli strain DH5alpha and Pseudomonas putida strain KT2440, the enzyme is exxpressed in both hosts but only active in Pseudomonas putida Pseudomonas sp.

Protein Variants

Protein Variants Comment Organism
H456R site-directed mutagenesis Pseudomonas sp.
additional information construction of the nox deletion mutant strain, that loses the ability to degrade nicotine, but not pseudooxynicotine Pseudomonas sp.

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
(S)-nicotine + acceptor + H2O Pseudomonas sp. via N-methylmyosmine, which then spontaneously hydrolyzes to pseudooxynicotine. The two enantiomers are degraded at approximately the same rate, indicating that NOX does not show chiral selectivity pseudooxynicotine + reduced acceptor
-
?
(S)-nicotine + acceptor + H2O Pseudomonas sp. HZN6 via N-methylmyosmine, which then spontaneously hydrolyzes to pseudooxynicotine. The two enantiomers are degraded at approximately the same rate, indicating that NOX does not show chiral selectivity pseudooxynicotine + reduced acceptor
-
?

Organism

Organism UniProt Comment Textmining
no activity in Escherichia coli strain DH5alpha
-
-
-
no activity in Pseudomonas putida strain KT2440
-
-
-
Pseudomonas sp. M4T5L3 gene nox
-
Pseudomonas sp. HZN6 M4T5L3 gene nox
-

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
(RS)-nicotine + acceptor + H2O via N-methylmyosmine, which then spontaneously hydrolyzes to pseudooxynicotine. The two enantiomers are degraded at approximately the same rate, indicating that NOX does not show chiral selectivity Pseudomonas sp. pseudooxynicotine + reduced acceptor
-
?
(RS)-nicotine + acceptor + H2O via N-methylmyosmine, which then spontaneously hydrolyzes to pseudooxynicotine. The two enantiomers are degraded at approximately the same rate, indicating that NOX does not show chiral selectivity Pseudomonas sp. HZN6 pseudooxynicotine + reduced acceptor
-
?
(S)-nicotine + acceptor + H2O via N-methylmyosmine, which then spontaneously hydrolyzes to pseudooxynicotine. The two enantiomers are degraded at approximately the same rate, indicating that NOX does not show chiral selectivity Pseudomonas sp. pseudooxynicotine + reduced acceptor
-
?
(S)-nicotine + acceptor + H2O via N-methylmyosmine, which then spontaneously hydrolyzes to pseudooxynicotine. The two enantiomers are degraded at approximately the same rate, indicating that NOX does not show chiral selectivity Pseudomonas sp. HZN6 pseudooxynicotine + reduced acceptor
-
?

Cofactor

Cofactor Comment Organism Structure
FAD conserved FAD-binding GXGXXG Pseudomonas sp.

General Information

General Information Comment Organism
malfunction a nox disruption mutant of strain HZN6 loses the ability to degrade nicotine, but not pseudooxynicotine Pseudomonas sp.
metabolism the enzyme is responsible for the first step of nicotine degradation Pseudomonas sp.
additional information the conserved FAD-binding GXGXXG motif and His456 are essential for nicotine degradation activity Pseudomonas sp.
physiological function the enzyme nonenantioselectively degrades nicotine to pseudooxynicotine Pseudomonas sp.