Information on EC 1.14.13.163 - 6-hydroxy-3-succinoylpyridine 3-monooxygenase

for references in articles please use BRENDA:EC1.14.13.163
Word Map on EC 1.14.13.163
Please wait a moment until all data is loaded. This message will disappear when all data is loaded.
Specify your search results
Select one or more organisms in this record:
Show additional data
Do not include text mining results
Include (text mining) results
Include results (AMENDA + additional results, but less precise)

The enzyme appears in viruses and cellular organisms

EC NUMBER
COMMENTARY hide
1.14.13.163
-
RECOMMENDED NAME
GeneOntology No.
6-hydroxy-3-succinoylpyridine 3-monooxygenase
-
REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
4-(6-hydroxypyridin-3-yl)-4-oxobutanoate + 2 NADH + 2 H+ + O2 = 2,5-dihydroxypyridine + succinate semialdehyde + 2 NAD+ + H2O
show the reaction diagram
PATHWAY
BRENDA Link
KEGG Link
MetaCyc Link
nicotine degradation II (pyrrolidine pathway)
-
-
nicotine degradation III (VPP pathway)
-
-
Nicotinate and nicotinamide metabolism
-
-
Microbial metabolism in diverse environments
-
-
SYSTEMATIC NAME
IUBMB Comments
4-(6-hydroxypyridin-3-yl)-4-oxobutanoate,NADH:oxygen oxidoreductase (3-hydroxylating, succinate semialdehyde releasing)
The enzyme catalyses a reaction in the nicotine degradation pathway of Pseudomonas species. One of the enzymes from the soil bacterium Pseudomonas putida S16 contains an FAD cofactor [2].
ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
isolated from soil samples obtained from a field under continuous tobacco cropping in Henan, P.R. China
-
-
Manually annotated by BRENDA team
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
evolution
metabolism
physiological function
additional information
SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
4-(6-hydroxypyridin-3-yl)-4-oxobutanoate + 2 NADH + 2 H+ + O2
2,5-dihydroxypyridine + succinate semialdehyde + 2 NAD+ + H2O
show the reaction diagram
6-hydroxy-3-succinoylpyridine + 2 NADH + 2 H+ + O2
2,5-dihydroxypyridine + succinate semialdehyde + 2 NAD+ + H2O
show the reaction diagram
additional information
?
-
NATURAL SUBSTRATES
NATURAL PRODUCTS
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
4-(6-hydroxypyridin-3-yl)-4-oxobutanoate + 2 NADH + 2 H+ + O2
2,5-dihydroxypyridine + succinate semialdehyde + 2 NAD+ + H2O
show the reaction diagram
COFACTOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
additional information
-
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
Cu2+
2 mM, complete loss of activity
Zn2+
2 mM, complete loss of activity
additional information
-
the enzyme is not significantly affected by Ni2+, Ca2+, K+, and Na+ at 5 mM. The enzyme is no metalloenzyme
INHIBITORS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
acetone
-
inhibits at 20% v/v
Cd2+
strong inhibition
Chloroform
-
inhibits at 20% v/v
Co2+
-
inhibits at 5 mM
dimethylformamide
-
inhibits at 20% v/v
DMSO
-
inhibits slightly by 10% at 20% v/v
EDTA
-
inhibits 15-20% at 0.5-5 mM
Fe2+
-
inhibits at 5 mM
formaldehyde
-
inhibits at 20% v/v
methanol
-
inhibits at 20% v/v
Mg2+
-
slight inhibition
Mn2+
-
slight inhibition
Span-20
-
at 1% w/v by over 50%
SPAN-40
-
at 1% w/v by over 50%
Toluene
-
inhibits at 20% v/v
Triton X-100
-
at 1% w/v by over 50%
additional information
-
no inhibition by Twen-20, Tween-40, and Tween-80 at 1% w/v
-
ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
6-hydroxy-3-succinoylpyridine
acts as an initial effector and strongly stimulates NADH oxidation
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.104 - 0.201
4-(6-hydroxypyridin-3-yl)-4-oxobutanoate
0.0294 - 0.23
NADH
additional information
additional information
-
stopped-flow spectroscopy and kinetic analysis
-
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
2.1 - 21.6
4-(6-hydroxypyridin-3-yl)-4-oxobutanoate
1.3 - 17.5
NADH
kcat/KM VALUE [1/mMs-1]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
11.7
4-(6-hydroxypyridin-3-yl)-4-oxobutanoate
-
pH 8.5, 30°C
5.7
NADH
-
pH 8.5, 30°C
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
4.8
pH 8.0, 25°C
5.1
-
purified native enzyme, pH 8.5, 30°C
10.3
purified native enzyme, pH 8.0, 30°C
18.8
purified recombinant enzyme, pH 8.0, 30°C
31.8
-
purified recombinant His-tagged enzyme, pH 8.5, 30°C
pH RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
6 - 9.7
-
activity range, profile overview
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
TEMPERATURE RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
5 - 45
-
activity range, profile overview
SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
SOURCE
strain SJY uses nicotine as a sole source of carbon, nitrogen, and energy
Manually annotated by BRENDA team
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
38000
x * 38000, SDS-PAGE and calculated
40000
2 * 40000, SDS-PAGE
90000
gel filtration
SUBUNITS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
homodimer
pH STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
25
30 min, stable up to
30
-
purified enzyme, pH 8.5, 120 min, only slight loss of activity
40
-
purified enzyme, pH 8.5, 30 min, loss of 60% activity
STORAGE STABILITY
ORGANISM
UNIPROT
LITERATURE
-20°C, Tris-HCl buffer, stable for several months
-80°C, purified recombinant His-tagged enzyme HspB, as a frozen solution or dry powder, no change in activity for at least 3 months, pH 8.0
-
4°C Tris-HCl buffer, stable for at least one week
Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
native enzyme 17.6fold to homogeneity by ammonium sulfate fractionation, anion exchange and hydrophobic interaction chromatography, and gel filtration, recombinant His-tagged enzyme 6.2fold from Escherichia coli strain B21-Codon Plus(DE3)-RIL by nickel affinity chromatography and gel filtration
-
native enzyme 64.4fold from Agrobacterium tumefaciens strain S33 cells by ammonium sulfate fractionation, hydrophobic interaction chromatography, desalting gel filtration, and two different steps of anion exchange chromatography, followed by gel filtration. Recombinant His-tagged enzyme from Escherichia coli strain BL21(DE3) by nickel affinity chromatography, anion exchange chromatography, and ultrafiltration. During purification, all the buffers are supplemented with 0.005 mM FAD. When FAD is omitted, the purification yield is very low
recombinant His-tagged enzyme from Escherichia coli strain BL21(DE3) by nickel affinity chromatography. FAD is partly lost during the purification
recombinant His6-tagged enzyme from Escherichia coli strain BL21(DE3) by nickel affinity chromatography
-
Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
DNA and amino acid sequence determination and analysis, sequence comparisons, RT-PCR expression analysis, recombinant expression of His-tagged enzyme in Escherichia coli strain BL21(DE3)
expression in Escherichia coli
gene hspB, phylogenetic analysis and tree, recombinant expression of His6-tagged enzyme in Escherichia coli strain BL21(DE3)
-
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)
recombinant expression of His-tagged enzyme in Escherichia coli strain B21-Codon Plus(DE3)-RIL
-
ENGINEERING
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
additional information
APPLICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
synthesis