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Literature summary for 1.5.3.5 extracted from

  • Qiu, J.; Wei, Y.; Ma, Y.; Wen, R.; Wen, Y.; Liu, W.
    A novel (S)-6-hydroxynicotine oxidase gene from Shinella sp. strain HZN7 (2014), Appl. Environ. Microbiol., 80, 5552-5560 .
    View publication on PubMedView publication on EuropePMC

Cloned(Commentary)

Cloned (Comment) Organism
overexpressed in Escherichia coli BL21(DE3) as a C-terminal His6-tagged fusion protein Shinella sp.

Inhibitors

Inhibitors Comment Organism Structure
Ag+ 1 mM, about 85% inhibition Shinella sp.
Co2+ 1 mM, about 10% inhibition Shinella sp.
Cu2+ 1 mM, about 85% inhibition Shinella sp.
EDTA incubation for 1 h completely inhibits enzymatic activity Shinella sp.
Hg2+ 1 mM, about 80% inhibition Shinella sp.
SDS incubation for 1 h completely inhibits enzymatic activity Shinella sp.
Zn2+ 1 mM, about 25% inhibition Shinella sp.

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.019
-
(S)-6-hydroxynicotine pH 7.0, 40┬░C Shinella sp.
0.396
-
(S)-nicotine pH 7.0, 40┬░C Shinella sp.

Metals/Ions

Metals/Ions Comment Organism Structure
additional information the enzyme does not require Al3+, Ca2+, Mg2+, or Mn2+ for its activity Shinella sp.

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
(S)-6-hydroxynicotine + H2O + O2 Shinella sp. the enzyme is essential for nicotine or (S)-6-hydroxynicotine degradation 1-(6-hydroxypyridin-3-yl)-4-(methylamino)butan-1-one + H2O2
-
?

Organism

Organism UniProt Comment Textmining
Shinella sp. A0A075BSX9
-
-

Purification (Commentary)

Purification (Comment) Organism
-
Shinella sp.

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
(S)-6-hydroxynicotine + H2O + O2 the enzyme is essential for nicotine or (S)-6-hydroxynicotine degradation Shinella sp. 1-(6-hydroxypyridin-3-yl)-4-(methylamino)butan-1-one + H2O2
-
?
(S)-6-hydroxynicotine + H2O + O2 the enzyme converts (S)-6-hydroxynicotine into 6-hydroxy-N-methylmyosmine which then spontaneously hydrolyzes into 6-hydroxypseudooxynicotine. (S)-6-Hydroxynicotine is the preferred substrate in vivo. The enzyme shows no activities toward the R enantiomer of nicotine or 6-hydroxynicotine Shinella sp. 1-(6-hydroxypyridin-3-yl)-4-(methylamino)butan-1-one + H2O2
-
?
(S)-nicotine + H2O + O2 the enzyme converts (S)-nicotine into N-methylmyosmine, which spontaneously hydrolyzes into pseudooxynicotine Shinella sp. N-methylmyosmine + H2O2
-
?

Subunits

Subunits Comment Organism
homodimer 2 * 50000, SDS-PAGE Shinella sp.

Temperature Optimum [┬░C]

Temperature Optimum [┬░C] Temperature Optimum Maximum [┬░C] Comment Organism
40
-
assay at Shinella sp.

Temperature Range [┬░C]

Temperature Minimum [┬░C] Temperature Maximum [┬░C] Comment Organism
20 50 20┬░C: about 65% of maximal activity, 50┬░C: about 70% of maximal activity Shinella sp.

Temperature Stability [┬░C]

Temperature Stability Minimum [┬░C] Temperature Stability Maximum [┬░C] Comment Organism
5 40 1 h, no loss of activity Shinella sp.
50
-
1 h, about 15% loss of activity Shinella sp.
60
-
1 h, about 65% loss of activity Shinella sp.

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
2.03
-
(S)-nicotine pH 7.0, 40┬░C Shinella sp.
7.3
-
(S)-6-hydroxynicotine pH 7.0, 40┬░C Shinella sp.

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
7
-
-
Shinella sp.

pH Stability

pH Stability pH Stability Maximum Comment Organism
5
-
40┬░C, 1 h, about 50% loss of activity Shinella sp.
7 8 40┬░C, 1 h, no loss of activity Shinella sp.
10
-
40┬░C, 1 h, about 30% loss of activity Shinella sp.

Cofactor

Cofactor Comment Organism Structure
FAD flavoprotein. FAD content is determined to be 1.0 of FAD per mole subunit Shinella sp.

General Information

General Information Comment Organism
metabolism the deletion and complementation of the nctB gene shows that this enzyme is essential for nicotine or (S)-6-hydroxynicotine degradation Shinella sp.

kcat/KM [mM/s]

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
0.195
-
(S)-nicotine pH 7.0, 40┬░C Shinella sp.
379
-
(S)-6-hydroxynicotine pH 7.0, 40┬░C Shinella sp.