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Literature summary 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

Application

EC Number Application Comment Organism
1.14.13.163 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)

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

Inhibitors

EC Number Inhibitors Comment Organism Structure
1.14.13.163 acetone inhibits at 20% v/v Pseudomonas sp. ZZ-5
1.14.13.163 chloroform inhibits at 20% v/v Pseudomonas sp. ZZ-5
1.14.13.163 Co2+ inhibits at 5 mM Pseudomonas sp. ZZ-5
1.14.13.163 Cu2+ inhibits at 5 mM Pseudomonas sp. ZZ-5
1.14.13.163 dimethylformamide inhibits at 20% v/v Pseudomonas sp. ZZ-5
1.14.13.163 DMSO inhibits slightly by 10% at 20% v/v Pseudomonas sp. ZZ-5
1.14.13.163 EDTA inhibits 15-20% at 0.5-5 mM Pseudomonas sp. ZZ-5
1.14.13.163 Fe2+ inhibits at 5 mM Pseudomonas sp. ZZ-5
1.14.13.163 formaldehyde inhibits at 20% v/v Pseudomonas sp. ZZ-5
1.14.13.163 methanol inhibits at 20% v/v Pseudomonas sp. ZZ-5
1.14.13.163 Mg2+ slight inhibition Pseudomonas sp. ZZ-5
1.14.13.163 Mn2+ slight inhibition Pseudomonas sp. ZZ-5
1.14.13.163 additional information no inhibition by Twen-20, Tween-40, and Tween-80 at 1% w/v Pseudomonas sp. ZZ-5
1.14.13.163 Span-20 at 1% w/v by over 50% Pseudomonas sp. ZZ-5
1.14.13.163 SPAN-40 at 1% w/v by over 50% Pseudomonas sp. ZZ-5
1.14.13.163 Toluene inhibits at 20% v/v Pseudomonas sp. ZZ-5
1.14.13.163 Triton X-100 at 1% w/v by over 50% Pseudomonas sp. ZZ-5
1.14.13.163 Zn2+ inhibits at 5 mM Pseudomonas sp. ZZ-5

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
1.14.13.163 0.18
-
4-(6-hydroxypyridin-3-yl)-4-oxobutanoate pH 8.5, 30°C Pseudomonas sp. ZZ-5
1.14.13.163 0.23
-
NADH pH 8.5, 30°C Pseudomonas sp. ZZ-5

Metals/Ions

EC Number Metals/Ions Comment Organism Structure
1.14.13.163 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)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
1.14.13.163 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

EC Number Organism UniProt Comment Textmining
1.14.13.163 Pseudomonas sp. ZZ-5
-
isolated from soil samples obtained from a field under continuous tobacco cropping in Henan, P.R. China
-

Purification (Commentary)

EC Number Purification (Comment) Organism
1.14.13.163 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]

EC Number Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
1.14.13.163 5.1
-
purified native enzyme, pH 8.5, 30°C Pseudomonas sp. ZZ-5
1.14.13.163 31.8
-
purified recombinant His-tagged enzyme, pH 8.5, 30°C Pseudomonas sp. ZZ-5

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
1.14.13.163 4-(6-hydroxypyridin-3-yl)-4-oxobutanoate + 2 NADH + 2 H+ + O2
-
Pseudomonas sp. ZZ-5 2,5-dihydroxypyridine + succinate semialdehyde + 2 NAD+ + H2O
-
?
1.14.13.163 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
-
?
1.14.13.163 additional information assay method optimization, overview. Product identification by LC-MS Pseudomonas sp. ZZ-5 ?
-
?

Subunits

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

Synonyms

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

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
1.14.13.163 30
-
-
Pseudomonas sp. ZZ-5

Temperature Range [°C]

EC Number Temperature Minimum [°C] Temperature Maximum [°C] Comment Organism
1.14.13.163 5 45 activity range, profile overview Pseudomonas sp. ZZ-5

Temperature Stability [°C]

EC Number Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
1.14.13.163 30
-
purified enzyme, pH 8.5, 120 min, only slight loss of activity Pseudomonas sp. ZZ-5
1.14.13.163 40
-
purified enzyme, pH 8.5, 30 min, loss of 60% activity Pseudomonas sp. ZZ-5

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
1.14.13.163 1.3
-
NADH pH 8.5, 30°C Pseudomonas sp. ZZ-5
1.14.13.163 2.1
-
4-(6-hydroxypyridin-3-yl)-4-oxobutanoate pH 8.5, 30°C Pseudomonas sp. ZZ-5

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
1.14.13.163 8.5
-
-
Pseudomonas sp. ZZ-5

pH Range

EC Number pH Minimum pH Maximum Comment Organism
1.14.13.163 6 9.7 activity range, profile overview Pseudomonas sp. ZZ-5

pH Stability

EC Number pH Stability pH Stability Maximum Comment Organism
1.14.13.163 7 9 purified enzyme, 30°C, 30 min, over 50% activity remaining within this range Pseudomonas sp. ZZ-5

Cofactor

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

kcat/KM [mM/s]

EC Number kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
1.14.13.163 5.7
-
NADH pH 8.5, 30°C Pseudomonas sp. ZZ-5
1.14.13.163 11.7
-
4-(6-hydroxypyridin-3-yl)-4-oxobutanoate pH 8.5, 30°C Pseudomonas sp. ZZ-5