BRENDA - Enzyme Database show
show all sequences of 1.14.13.163

Characterization of a novel nicotine hydroxylase from Pseudomonas sp. ZZ-5 that catalyzes the conversion of 6-hydroxy-3-succinoylpyridine into 2,5-dihydroxypyridine

Wei, T.; Zang, J.; Zheng, Y.; Tang, H.; Huang, S.; Mao, D.; Catalysts 7, 272-281 (2017)

Data extracted from this reference:

Application
Application
Commentary
Organism
synthesis
enzyme HSPHZZ can be used for the enzymatic production of 2,5-dihydroxypyridine in biotechnology applications. 85.3 mg/l 2,5-dihydroxypyridine is produced in 40 min with a conversion of 74.9% at 30C, pH 8.5, 1.0 mM substrate concentration, and 0.001 mM enzyme concentration
Pseudomonas sp. ZZ-5
Cloned(Commentary)
Commentary
Organism
recombinant expression of His-tagged enzyme in Escherichia coli strain B21-Codon Plus(DE3)-RIL
Pseudomonas sp. ZZ-5
Inhibitors
Inhibitors
Commentary
Organism
Structure
acetone
inhibits at 20% v/v
Pseudomonas sp. ZZ-5
Chloroform
inhibits at 20% v/v
Pseudomonas sp. ZZ-5
Co2+
inhibits at 5 mM
Pseudomonas sp. ZZ-5
Cu2+
inhibits at 5 mM
Pseudomonas sp. ZZ-5
dimethylformamide
inhibits at 20% v/v
Pseudomonas sp. ZZ-5
DMSO
inhibits slightly by 10% at 20% v/v
Pseudomonas sp. ZZ-5
EDTA
inhibits 15-20% at 0.5-5 mM
Pseudomonas sp. ZZ-5
Fe2+
inhibits at 5 mM
Pseudomonas sp. ZZ-5
formaldehyde
inhibits at 20% v/v
Pseudomonas sp. ZZ-5
methanol
inhibits at 20% v/v
Pseudomonas sp. ZZ-5
Mg2+
slight inhibition
Pseudomonas sp. ZZ-5
Mn2+
slight inhibition
Pseudomonas sp. ZZ-5
additional information
no inhibition by Twen-20, Tween-40, and Tween-80 at 1% w/v
Pseudomonas sp. ZZ-5
Span-20
at 1% w/v by over 50%
Pseudomonas sp. ZZ-5
SPAN-40
at 1% w/v by over 50%
Pseudomonas sp. ZZ-5
Toluene
inhibits at 20% v/v
Pseudomonas sp. ZZ-5
Triton X-100
at 1% w/v by over 50%
Pseudomonas sp. ZZ-5
Zn2+
inhibits at 5 mM
Pseudomonas sp. ZZ-5
KM Value [mM]
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
0.18
-
4-(6-hydroxypyridin-3-yl)-4-oxobutanoate
pH 8.5, 30C
Pseudomonas sp. ZZ-5
0.23
-
NADH
pH 8.5, 30C
Pseudomonas sp. ZZ-5
Metals/Ions
Metals/Ions
Commentary
Organism
Structure
additional information
the enzyme is not significantly affected by Ni2+, Ca2+, K+, and Na+ at 5 mM. The enzyme is no metalloenzyme
Pseudomonas sp. ZZ-5
Natural Substrates/ Products (Substrates)
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
4-(6-hydroxypyridin-3-yl)-4-oxobutanoate + 2 NADH + 2 H+ + O2
Pseudomonas sp. ZZ-5
-
2,5-dihydroxypyridine + succinate semialdehyde + 2 NAD+ + H2O
-
-
?
Organism
Organism
Primary Accession No. (UniProt)
Commentary
Textmining
Pseudomonas sp. ZZ-5
-
isolated from soil samples obtained from a field under continuous tobacco cropping in Henan, P.R. China
-
Purification (Commentary)
Commentary
Organism
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
Pseudomonas sp. ZZ-5
Specific Activity [micromol/min/mg]
Specific Activity Minimum [mol/min/mg]
Specific Activity Maximum [mol/min/mg]
Commentary
Organism
5.1
-
purified native enzyme, pH 8.5, 30C
Pseudomonas sp. ZZ-5
31.8
-
purified recombinant His-tagged enzyme, pH 8.5, 30C
Pseudomonas sp. ZZ-5
Substrates and Products (Substrate)
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
4-(6-hydroxypyridin-3-yl)-4-oxobutanoate + 2 NADH + 2 H+ + O2
-
744640
Pseudomonas sp. ZZ-5
2,5-dihydroxypyridine + succinate semialdehyde + 2 NAD+ + H2O
-
-
-
?
4-(6-hydroxypyridin-3-yl)-4-oxobutanoate + 2 NADH + 2 H+ + O2
the enzyme efficiently catalyzes the conversion of 6-hydroxy-3-succinoylpyridine (HSP) into 2,5-dihydroxypyridine (2,5-DHP) and succinic acid in the presence of NADH and FAD
744640
Pseudomonas sp. ZZ-5
2,5-dihydroxypyridine + succinate semialdehyde + 2 NAD+ + H2O
-
-
-
?
additional information
assay method optimization, overview. Product identification by LC-MS
744640
Pseudomonas sp. ZZ-5
?
-
-
-
-
Subunits
Subunits
Commentary
Organism
?
x * 45000, SDS-PAGE, x * 44300 , about, sequence calculation
Pseudomonas sp. ZZ-5
Temperature Optimum [C]
Temperature Optimum [C]
Temperature Optimum Maximum [C]
Commentary
Organism
30
-
-
Pseudomonas sp. ZZ-5
Temperature Range [C]
Temperature Minimum [C]
Temperature Maximum [C]
Commentary
Organism
5
45
activity range, profile overview
Pseudomonas sp. ZZ-5
Temperature Stability [C]
Temperature Stability Minimum [C]
Temperature Stability Maximum [C]
Commentary
Organism
30
-
purified enzyme, pH 8.5, 120 min, only slight loss of activity
Pseudomonas sp. ZZ-5
40
-
purified enzyme, pH 8.5, 30 min, loss of 60% activity
Pseudomonas sp. ZZ-5
Turnover Number [1/s]
Turnover Number Minimum [1/s]
Turnover Number Maximum [1/s]
Substrate
Commentary
Organism
Structure
1.3
-
NADH
pH 8.5, 30C
Pseudomonas sp. ZZ-5
2.1
-
4-(6-hydroxypyridin-3-yl)-4-oxobutanoate
pH 8.5, 30C
Pseudomonas sp. ZZ-5
pH Optimum
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
8.5
-
-
Pseudomonas sp. ZZ-5
pH Range
pH Minimum
pH Maximum
Commentary
Organism
6
9.7
activity range, profile overview
Pseudomonas sp. ZZ-5
pH Stability
pH Stability
pH Stability Maximum
Commentary
Organism
7
9
purified enzyme, 30C, 30 min, over 50% activity remaining within this range
Pseudomonas sp. ZZ-5
Cofactor
Cofactor
Commentary
Organism
Structure
FAD
-
Pseudomonas sp. ZZ-5
NADH
-
Pseudomonas sp. ZZ-5
Application (protein specific)
Application
Commentary
Organism
synthesis
enzyme HSPHZZ can be used for the enzymatic production of 2,5-dihydroxypyridine in biotechnology applications. 85.3 mg/l 2,5-dihydroxypyridine is produced in 40 min with a conversion of 74.9% at 30C, pH 8.5, 1.0 mM substrate concentration, and 0.001 mM enzyme concentration
Pseudomonas sp. ZZ-5
Cloned(Commentary) (protein specific)
Commentary
Organism
recombinant expression of His-tagged enzyme in Escherichia coli strain B21-Codon Plus(DE3)-RIL
Pseudomonas sp. ZZ-5
Cofactor (protein specific)
Cofactor
Commentary
Organism
Structure
FAD
-
Pseudomonas sp. ZZ-5
NADH
-
Pseudomonas sp. ZZ-5
Inhibitors (protein specific)
Inhibitors
Commentary
Organism
Structure
acetone
inhibits at 20% v/v
Pseudomonas sp. ZZ-5
Chloroform
inhibits at 20% v/v
Pseudomonas sp. ZZ-5
Co2+
inhibits at 5 mM
Pseudomonas sp. ZZ-5
Cu2+
inhibits at 5 mM
Pseudomonas sp. ZZ-5
dimethylformamide
inhibits at 20% v/v
Pseudomonas sp. ZZ-5
DMSO
inhibits slightly by 10% at 20% v/v
Pseudomonas sp. ZZ-5
EDTA
inhibits 15-20% at 0.5-5 mM
Pseudomonas sp. ZZ-5
Fe2+
inhibits at 5 mM
Pseudomonas sp. ZZ-5
formaldehyde
inhibits at 20% v/v
Pseudomonas sp. ZZ-5
methanol
inhibits at 20% v/v
Pseudomonas sp. ZZ-5
Mg2+
slight inhibition
Pseudomonas sp. ZZ-5
Mn2+
slight inhibition
Pseudomonas sp. ZZ-5
additional information
no inhibition by Twen-20, Tween-40, and Tween-80 at 1% w/v
Pseudomonas sp. ZZ-5
Span-20
at 1% w/v by over 50%
Pseudomonas sp. ZZ-5
SPAN-40
at 1% w/v by over 50%
Pseudomonas sp. ZZ-5
Toluene
inhibits at 20% v/v
Pseudomonas sp. ZZ-5
Triton X-100
at 1% w/v by over 50%
Pseudomonas sp. ZZ-5
Zn2+
inhibits at 5 mM
Pseudomonas sp. ZZ-5
KM Value [mM] (protein specific)
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
0.18
-
4-(6-hydroxypyridin-3-yl)-4-oxobutanoate
pH 8.5, 30C
Pseudomonas sp. ZZ-5
0.23
-
NADH
pH 8.5, 30C
Pseudomonas sp. ZZ-5
Metals/Ions (protein specific)
Metals/Ions
Commentary
Organism
Structure
additional information
the enzyme is not significantly affected by Ni2+, Ca2+, K+, and Na+ at 5 mM. The enzyme is no metalloenzyme
Pseudomonas sp. ZZ-5
Natural Substrates/ Products (Substrates) (protein specific)
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
4-(6-hydroxypyridin-3-yl)-4-oxobutanoate + 2 NADH + 2 H+ + O2
Pseudomonas sp. ZZ-5
-
2,5-dihydroxypyridine + succinate semialdehyde + 2 NAD+ + H2O
-
-
?
Purification (Commentary) (protein specific)
Commentary
Organism
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
Pseudomonas sp. ZZ-5
Specific Activity [micromol/min/mg] (protein specific)
Specific Activity Minimum [mol/min/mg]
Specific Activity Maximum [mol/min/mg]
Commentary
Organism
5.1
-
purified native enzyme, pH 8.5, 30C
Pseudomonas sp. ZZ-5
31.8
-
purified recombinant His-tagged enzyme, pH 8.5, 30C
Pseudomonas sp. ZZ-5
Substrates and Products (Substrate) (protein specific)
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
4-(6-hydroxypyridin-3-yl)-4-oxobutanoate + 2 NADH + 2 H+ + O2
-
744640
Pseudomonas sp. ZZ-5
2,5-dihydroxypyridine + succinate semialdehyde + 2 NAD+ + H2O
-
-
-
?
4-(6-hydroxypyridin-3-yl)-4-oxobutanoate + 2 NADH + 2 H+ + O2
the enzyme efficiently catalyzes the conversion of 6-hydroxy-3-succinoylpyridine (HSP) into 2,5-dihydroxypyridine (2,5-DHP) and succinic acid in the presence of NADH and FAD
744640
Pseudomonas sp. ZZ-5
2,5-dihydroxypyridine + succinate semialdehyde + 2 NAD+ + H2O
-
-
-
?
additional information
assay method optimization, overview. Product identification by LC-MS
744640
Pseudomonas sp. ZZ-5
?
-
-
-
-
Subunits (protein specific)
Subunits
Commentary
Organism
?
x * 45000, SDS-PAGE, x * 44300 , about, sequence calculation
Pseudomonas sp. ZZ-5
Temperature Optimum [C] (protein specific)
Temperature Optimum [C]
Temperature Optimum Maximum [C]
Commentary
Organism
30
-
-
Pseudomonas sp. ZZ-5
Temperature Range [C] (protein specific)
Temperature Minimum [C]
Temperature Maximum [C]
Commentary
Organism
5
45
activity range, profile overview
Pseudomonas sp. ZZ-5
Temperature Stability [C] (protein specific)
Temperature Stability Minimum [C]
Temperature Stability Maximum [C]
Commentary
Organism
30
-
purified enzyme, pH 8.5, 120 min, only slight loss of activity
Pseudomonas sp. ZZ-5
40
-
purified enzyme, pH 8.5, 30 min, loss of 60% activity
Pseudomonas sp. ZZ-5
Turnover Number [1/s] (protein specific)
Turnover Number Minimum [1/s]
Turnover Number Maximum [1/s]
Substrate
Commentary
Organism
Structure
1.3
-
NADH
pH 8.5, 30C
Pseudomonas sp. ZZ-5
2.1
-
4-(6-hydroxypyridin-3-yl)-4-oxobutanoate
pH 8.5, 30C
Pseudomonas sp. ZZ-5
pH Optimum (protein specific)
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
8.5
-
-
Pseudomonas sp. ZZ-5
pH Range (protein specific)
pH Minimum
pH Maximum
Commentary
Organism
6
9.7
activity range, profile overview
Pseudomonas sp. ZZ-5
pH Stability (protein specific)
pH Stability
pH Stability Maximum
Commentary
Organism
7
9
purified enzyme, 30C, 30 min, over 50% activity remaining within this range
Pseudomonas sp. ZZ-5
KCat/KM [mM/s]
kcat/KM Value [1/mMs-1]
kcat/KM Value Maximum [1/mMs-1]
Substrate
Commentary
Organism
Structure
5.7
-
NADH
pH 8.5, 30C
Pseudomonas sp. ZZ-5
11.7
-
4-(6-hydroxypyridin-3-yl)-4-oxobutanoate
pH 8.5, 30C
Pseudomonas sp. ZZ-5
KCat/KM [mM/s] (protein specific)
KCat/KM Value [1/mMs-1]
KCat/KM Value Maximum [1/mMs-1]
Substrate
Commentary
Organism
Structure
5.7
-
NADH
pH 8.5, 30C
Pseudomonas sp. ZZ-5
11.7
-
4-(6-hydroxypyridin-3-yl)-4-oxobutanoate
pH 8.5, 30C
Pseudomonas sp. ZZ-5
Other publictions for EC 1.14.13.163
No.
1st author
Pub Med
title
organims
journal
volume
pages
year
Activating Compound
Application
Cloned(Commentary)
Crystallization (Commentary)
Engineering
General Stability
Inhibitors
KM Value [mM]
Localization
Metals/Ions
Molecular Weight [Da]
Natural Substrates/ Products (Substrates)
Organic Solvent Stability
Organism
Oxidation Stability
Posttranslational Modification
Purification (Commentary)
Reaction
Renatured (Commentary)
Source Tissue
Specific Activity [micromol/min/mg]
Storage Stability
Substrates and Products (Substrate)
Subunits
Temperature Optimum [C]
Temperature Range [C]
Temperature Stability [C]
Turnover Number [1/s]
pH Optimum
pH Range
pH Stability
Cofactor
Ki Value [mM]
pI Value
IC50 Value
Activating Compound (protein specific)
Application (protein specific)
Cloned(Commentary) (protein specific)
Cofactor (protein specific)
Crystallization (Commentary) (protein specific)
Engineering (protein specific)
General Stability (protein specific)
IC50 Value (protein specific)
Inhibitors (protein specific)
Ki Value [mM] (protein specific)
KM Value [mM] (protein specific)
Localization (protein specific)
Metals/Ions (protein specific)
Molecular Weight [Da] (protein specific)
Natural Substrates/ Products (Substrates) (protein specific)
Organic Solvent Stability (protein specific)
Oxidation Stability (protein specific)
Posttranslational Modification (protein specific)
Purification (Commentary) (protein specific)
Renatured (Commentary) (protein specific)
Source Tissue (protein specific)
Specific Activity [micromol/min/mg] (protein specific)
Storage Stability (protein specific)
Substrates and Products (Substrate) (protein specific)
Subunits (protein specific)
Temperature Optimum [C] (protein specific)
Temperature Range [C] (protein specific)
Temperature Stability [C] (protein specific)
Turnover Number [1/s] (protein specific)
pH Optimum (protein specific)
pH Range (protein specific)
pH Stability (protein specific)
pI Value (protein specific)
Expression
General Information
General Information (protein specific)
Expression (protein specific)
KCat/KM [mM/s]
KCat/KM [mM/s] (protein specific)
744640
Wei
Characterization of a novel n ...
Pseudomonas sp. ZZ-5
Catalysts
7
272-281
2017
-
1
1
-
-
-
18
2
-
1
-
1
-
1
-
-
1
-
-
-
2
-
3
1
1
1
2
2
1
1
1
2
-
-
-
-
1
1
2
-
-
-
-
18
-
2
-
1
-
1
-
-
-
1
-
-
2
-
3
1
1
1
2
2
1
1
1
-
-
-
-
-
2
2
741698
Li
Nicotine dehydrogenase comple ...
Agrobacterium tumefaciens, Agrobacterium tumefaciens S33
Appl. Environ. Microbiol.
82
1745-1755
2016
-
-
-
-
-
-
-
-
-
-
-
2
-
3
-
-
-
-
-
-
-
-
2
-
-
-
-
-
-
-
-
2
-
-
-
-
-
-
2
-
-
-
-
-
-
-
-
-
-
2
-
-
-
-
-
-
-
-
2
-
-
-
-
-
-
-
-
-
-
2
2
-
-
-
741691
Yu
Molecular mechanism of nicoti ...
Ochrobactrum sp.
Appl. Environ. Microbiol.
81
272-281
2015
-
-
1
-
-
-
3
2
-
-
-
1
-
1
-
-
1
-
-
1
-
-
1
-
1
-
-
2
1
-
-
2
-
-
-
-
-
1
2
-
-
-
-
3
-
2
-
-
-
1
-
-
-
1
-
1
-
-
1
-
1
-
-
2
1
-
-
-
-
3
3
-
-
-
745310
Yu
Mechanism of the 6-hydroxy-3- ...
Pseudomonas putida, Pseudomonas putida S16
J. Biol. Chem.
289
29158-29170
2014
-
-
1
-
-
-
-
3
-
-
-
2
-
4
-
-
1
1
-
-
-
1
4
-
1
-
-
2
1
-
-
2
-
-
-
-
-
1
2
-
-
-
-
-
-
3
-
-
-
2
-
-
-
1
-
-
-
1
4
-
1
-
-
2
1
-
-
-
-
3
3
-
-
-
746248
Li
6-hydroxy-3-succinoylpyridine ...
Agrobacterium tumefaciens, Agrobacterium tumefaciens S33
PLoS ONE
9
e103324
2014
-
1
1
-
-
-
-
-
-
-
1
2
-
7
-
-
1
-
-
-
2
-
6
1
1
-
-
-
1
-
-
2
-
-
-
-
1
1
2
-
-
-
-
-
-
-
-
-
1
2
-
-
-
1
-
-
2
-
6
1
1
-
-
-
1
-
-
-
-
2
2
-
-
-
718677
Wang
Identification of nicotine bio ...
Agrobacterium tumefaciens, Agrobacterium tumefaciens S33
Appl. Microbiol. Biotechnol.
95
1567-1578
2012
-
-
-
-
-
-
-
-
-
-
-
-
-
9
-
-
-
-
-
-
-
-
2
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
2
-
-
-
-
-
-
-
-
-
-
1
1
-
-
-
719982
Tang
A novel NADH-dependent and FAD ...
Pseudomonas putida, Pseudomonas putida S16
J. Biol. Chem.
286
39179-39187
2011
1
-
-
-
1
-
-
2
-
2
2
-
-
9
-
-
1
-
-
-
1
2
2
1
1
-
1
1
1
-
1
3
-
-
-
1
-
-
3
-
1
-
-
-
-
2
-
2
2
-
-
-
-
1
-
-
1
2
2
1
1
-
1
1
1
-
1
-
-
1
1
-
-
-
690527
Tang
A novel gene, encoding 6-hydro ...
Pseudomonas putida, Pseudomonas putida S16
Appl. Environ. Microbiol.
74
1567-1574
2008
-
-
1
-
-
-
-
-
-
-
1
-
-
5
-
-
-
-
-
-
-
-
2
1
1
-
-
-
1
-
-
1
-
-
-
-
-
1
1
-
-
-
-
-
-
-
-
-
1
-
-
-
-
-
-
-
-
-
2
1
1
-
-
-
1
-
-
-
-
-
-
-
-
-