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Literature summary extracted from

  • Yu, H.; Tang, H.; Zhu, X.; Li, Y.; Xu, P.
    Molecular mechanism of nicotine degradation by a newly isolated strain, Ochrobactrum sp. strain SJY1 (2015), Appl. Environ. Microbiol., 81, 272-281 .
    View publication on PubMedView publication on EuropePMC

Cloned(Commentary)

EC Number Cloned (Comment) Organism
1.5.3.5 expression in Escherichia coli BL21(DE3) Ochrobactrum sp. SJY1
1.14.13.163 gene vppD cloned from a 97-kbp DNA fragment containing six nicotine degradation-related genes which is obtained by gap closing from the genome sequence of strain SJY1, DNA and amino acid sequence determination and analysis, sequence comparisons and phylogenetic analysis, recombinant expression of His-tagged enzyme in Escherichia coli strain BL21(DE3) Ochrobactrum sp.

Inhibitors

EC Number Inhibitors Comment Organism Structure
1.14.13.163 Cd2+ strong inhibition Ochrobactrum sp.
1.14.13.163 Cu2+ strong inhibition Ochrobactrum sp.
1.14.13.163 Zn2+ strong inhibition Ochrobactrum sp.

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
1.5.3.5 24.6
-
 (S)-6-hydroxynicotine pH 8.0, 25°C Ochrobactrum sp. SJY1
1.14.13.163 0.112
-
 NADH pH 8.0, 25°C, recombinant His-tagged enzyme Ochrobactrum sp.
1.14.13.163 0.201
-
 4-(6-hydroxypyridin-3-yl)-4-oxobutanoate pH 8.0, 25°C, recombinant His-tagged enzyme Ochrobactrum sp.

Metals/Ions

EC Number Metals/Ions Comment Organism Structure
1.5.3.5 additional information the reaction progresses without the participation of metal ions Ochrobactrum sp. SJY1

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
1.5.3.5 (S)-6-hydroxynicotine + H2O + O2 Ochrobactrum sp. SJY1 the enzyme is involved in degradation of nicotine 1-(6-hydroxypyridin-3-yl)-4-(methylamino)butan-1-one + H2O2
-
 ?
1.14.13.163 4-(6-hydroxypyridin-3-yl)-4-oxobutanoate + 2 NADH + 2 H+ + O2 Ochrobactrum sp.
-
 2,5-dihydroxypyridine + succinate semialdehyde + 2 NAD+ + H2O
-
 ?

Organism

EC Number Organism UniProt Comment Textmining
1.5.3.5 Ochrobactrum sp. SJY1 A0A075XFI8
-
-
1.14.13.163 Ochrobactrum sp. A0A075XAG5
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
1.5.3.5
-
 Ochrobactrum sp. SJY1
1.14.13.163 recombinant His-tagged enzyme from Escherichia coli strain BL21(DE3) by nickel affinity chromatography. FAD is partly lost during the purification Ochrobactrum sp.

Source Tissue

EC Number Source Tissue Comment Organism Textmining
1.14.13.163 culture condition:nicotinic acid-grown cell strain SJY uses nicotine as a sole source of carbon, nitrogen, and energy Ochrobactrum sp.
-

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
1.5.3.5 (S)-6-hydroxynicotine + H2O + O2
-
 Ochrobactrum sp. SJY1 1-(6-hydroxypyridin-3-yl)-4-(methylamino)butan-1-one + H2O2
-
 ?
1.5.3.5 (S)-6-hydroxynicotine + H2O + O2 the enzyme is involved in degradation of nicotine Ochrobactrum sp. SJY1 1-(6-hydroxypyridin-3-yl)-4-(methylamino)butan-1-one + H2O2
-
 ?
1.14.13.163 4-(6-hydroxypyridin-3-yl)-4-oxobutanoate + 2 NADH + 2 H+ + O2
-
 Ochrobactrum sp. 2,5-dihydroxypyridine + succinate semialdehyde + 2 NAD+ + H2O
-
 ?

Synonyms

EC Number Synonyms Comment Organism
1.5.3.5 6-hydroxy-L-nicotine oxidase
-
 Ochrobactrum sp. SJY1
1.5.3.5 6HLNO
-
 Ochrobactrum sp. SJY1
1.5.3.5 VppB
-
 Ochrobactrum sp. SJY1
1.14.13.163 VppD
-
 Ochrobactrum sp.

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
1.5.3.5 45
-
-
 Ochrobactrum sp. SJY1
1.14.13.163 25
-
-
 Ochrobactrum sp.

Temperature Range [°C]

EC Number Temperature Minimum [°C] Temperature Maximum [°C] Comment Organism
1.5.3.5 15 65 15°C: about 40% of maximal activity, 65°C: about 55% of maximal activity Ochrobactrum sp. SJY1

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
1.5.3.5 25.8
-
 (S)-6-hydroxynicotine pH 8.0, 25°C Ochrobactrum sp. SJY1
1.14.13.163 17.5
-
 NADH pH 8.0, 25°C, recombinant His-tagged enzyme Ochrobactrum sp.
1.14.13.163 21.6
-
 4-(6-hydroxypyridin-3-yl)-4-oxobutanoate pH 8.0, 25°C, recombinant His-tagged enzyme Ochrobactrum sp.

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
1.5.3.5 8
-
-
 Ochrobactrum sp. SJY1
1.14.13.163 8
-
-
 Ochrobactrum sp.

pH Range

EC Number pH Minimum pH Maximum Comment Organism
1.5.3.5 7 9 pH 7.0: about 65% of maximal activity, pH 9.0: about 25% of maximal activity Ochrobactrum sp. SJY1

Cofactor

EC Number Cofactor Comment Organism Structure
1.5.3.5 FAD tightly bound to the enzyme Ochrobactrum sp. SJY1
1.14.13.163 FAD
-
 Ochrobactrum sp.
1.14.13.163 NADH
-
 Ochrobactrum sp.

General Information

EC Number General Information Comment Organism
1.14.13.163 evolution sequence alignment and phylogenetic analysis suggests that the VPP pathway, which evolved independently from nicotinic acid degradation, might have a closer relationship with the pyrrolidine pathway Ochrobactrum sp.
1.14.13.163 metabolism strain SJY1 efficiently degrades nicotine via a variant of the pyridine and pyrrolidine pathways (the VPP pathway), highlighting bacterial metabolic diversity in relation to nicotine degradation, a 97-kbp DNA fragment containing six nicotine degradation-related genes is obtained by gap closing from the genome sequence of strain SJY1, gene vppD gene encodes an NADH-dependent flavin-containing monooxygenase, which catalyzes the hydroxylation of 6-hydroxy-3-succinoylpyridine to 2,5-dihydroxypyridine. Nicotine degradation pathway in strain SJY1, detailed overview Ochrobactrum sp.
1.14.13.163 additional information the ativity of VppD is 10fold higher than the activity of the hydroxylase (HspB) from Pseudomonas putida strain S16 Ochrobactrum sp.