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Literature summary for 1.14.13.163 extracted from

  • Wei, T.; Zang, J.; Zheng, Y.; Tang, H.; Huang, S.; Mao, D.
    Characterization of a novel nicotine hydroxylase from Pseudomonas sp. ZZ-5 that catalyzes the conversion of 6-hydroxy-3-succinoylpyridine into 2,5-dihydroxypyridine (2017), Catalysts, 7, 272-281 .
    View publication on PubMedView publication on EuropePMC
Search for more literature for 1.14.13.163

Application

Application Comment 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 30°C, pH 8.5, 1.0 mM substrate concentration, and 0.001 mM enzyme concentration Pseudomonas sp. ZZ-5

Cloned(Commentary)

Cloned (Comment) Organism
recombinant expression of His-tagged enzyme in Escherichia coli strain B21-Codon Plus(DE3)-RIL Pseudomonas sp. ZZ-5

Inhibitors

Inhibitors Comment 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 Comment Organism Structure
0.18
-
 4-(6-hydroxypyridin-3-yl)-4-oxobutanoate pH 8.5, 30°C Pseudomonas sp. ZZ-5
0.23
-
 NADH pH 8.5, 30°C Pseudomonas sp. ZZ-5

Metals/Ions

Metals/Ions Comment 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 Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
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 UniProt Comment Textmining
Pseudomonas sp. ZZ-5
-
 isolated from soil samples obtained from a field under continuous tobacco cropping in Henan, P.R. China
-

Purification (Commentary)

Purification (Comment) 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 [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
5.1
-
 purified native enzyme, pH 8.5, 30°C Pseudomonas sp. ZZ-5
31.8
-
 purified recombinant His-tagged enzyme, pH 8.5, 30°C Pseudomonas sp. ZZ-5

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
4-(6-hydroxypyridin-3-yl)-4-oxobutanoate + 2 NADH + 2 H+ + O2
-
 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 Pseudomonas sp. ZZ-5 2,5-dihydroxypyridine + succinate semialdehyde + 2 NAD+ + H2O
-
 ?
additional information assay method optimization, overview. Product identification by LC-MS Pseudomonas sp. ZZ-5 ?
-
 ?

Subunits

Subunits Comment Organism
? x * 45000, SDS-PAGE, x * 44300 , about, sequence calculation Pseudomonas sp. ZZ-5

Synonyms

Synonyms Comment Organism
HSP hydroxylase
-
 Pseudomonas sp. ZZ-5
HSPHZZ
-
 Pseudomonas sp. ZZ-5
nicotine hydroxylase
-
 Pseudomonas sp. ZZ-5

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
30
-
-
 Pseudomonas sp. ZZ-5

Temperature Range [°C]

Temperature Minimum [°C] Temperature Maximum [°C] Comment Organism
5 45 activity range, profile overview Pseudomonas sp. ZZ-5

Temperature Stability [°C]

Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment 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 Comment Organism Structure
1.3
-
 NADH pH 8.5, 30°C Pseudomonas sp. ZZ-5
2.1
-
 4-(6-hydroxypyridin-3-yl)-4-oxobutanoate pH 8.5, 30°C Pseudomonas sp. ZZ-5

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
8.5
-
-
 Pseudomonas sp. ZZ-5

pH Range

pH Minimum pH Maximum Comment Organism
6 9.7 activity range, profile overview Pseudomonas sp. ZZ-5

pH Stability

pH Stability pH Stability Maximum Comment Organism
7 9 purified enzyme, 30°C, 30 min, over 50% activity remaining within this range Pseudomonas sp. ZZ-5

Cofactor

Cofactor Comment Organism Structure
FAD
-
 Pseudomonas sp. ZZ-5
NADH
-
 Pseudomonas sp. ZZ-5

kcat/KM [mM/s]

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
5.7
-
 NADH pH 8.5, 30°C Pseudomonas sp. ZZ-5
11.7
-
 4-(6-hydroxypyridin-3-yl)-4-oxobutanoate pH 8.5, 30°C Pseudomonas sp. ZZ-5