Any feedback?
Please rate this page
(literature.php)
(0/150)

BRENDA support

Literature summary for 1.14.13.163 extracted from

  • Yu, H.; Hausinger, R.; Tang, H.; Xu, P.
    Mechanism of the 6-hydroxy-3-succinoyl-pyridine 3-monooxygenase flavoprotein from Pseudomonas putida S16 (2014), J. Biol. Chem., 289, 29158-29170 .
    View publication on PubMedView publication on EuropePMC

Cloned(Commentary)

Cloned (Comment) Organism
gene hspB, phylogenetic analysis and tree, recombinant expression of His6-tagged enzyme in Escherichia coli strain BL21(DE3) Pseudomonas putida

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
additional information
-
additional information stopped-flow spectroscopy and kinetic analysis Pseudomonas putida
0.0294
-
NADH pH 8.0, 25°C, recombinant His-tagged enzyme Pseudomonas putida
0.173
-
4-(6-hydroxypyridin-3-yl)-4-oxobutanoate pH 8.0, 25°C, recombinant His-tagged enzyme Pseudomonas putida

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 putida
-
2,5-dihydroxypyridine + succinate semialdehyde + 2 NAD+ + H2O
-
?
4-(6-hydroxypyridin-3-yl)-4-oxobutanoate + 2 NADH + 2 H+ + O2 Pseudomonas putida S16
-
2,5-dihydroxypyridine + succinate semialdehyde + 2 NAD+ + H2O
-
?

Organism

Organism UniProt Comment Textmining
Pseudomonas putida
-
-
-
Pseudomonas putida S16
-
-
-

Purification (Commentary)

Purification (Comment) Organism
recombinant His6-tagged enzyme from Escherichia coli strain BL21(DE3) by nickel affinity chromatography Pseudomonas putida

Reaction

Reaction Comment Organism Reaction ID
4-(6-hydroxypyridin-3-yl)-4-oxobutanoate + 2 NADH + 2 H+ + O2 = 2,5-dihydroxypyridine + succinate semialdehyde + 2 NAD+ + H2O catalytic mechanism, overview. In contrast to conclusions reported previously, the second product of the HspB reaction is shown to be succinate, with isotope labeling experiments providing direct evidence that the newly introduced oxygen atom of succinate is derived from H2O. Reduced HspB reacts with oxygen to form a C(4a)-(hydro)peroxyflavin intermediate before it is converted to the oxidized flavoenzyme species. The formed C(4a)-hydroperoxyflavin intermediate reacts with HSP to form an intermediate that is hydrolyzed to the products 2,5-dihydroxypyridine and succinate Pseudomonas putida

Storage Stability

Storage Stability Organism
-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 Pseudomonas putida

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 putida 2,5-dihydroxypyridine + succinate semialdehyde + 2 NAD+ + H2O
-
?
4-(6-hydroxypyridin-3-yl)-4-oxobutanoate + 2 NADH + 2 H+ + O2
-
Pseudomonas putida S16 2,5-dihydroxypyridine + succinate semialdehyde + 2 NAD+ + H2O
-
?
additional information mass spectrometric analysis of substrates and products, with recombinant enzyme Pseudomonas putida ?
-
?
additional information mass spectrometric analysis of substrates and products, with recombinant enzyme Pseudomonas putida S16 ?
-
?

Synonyms

Synonyms Comment Organism
6-hydroxy-3-succinoyl-pyridine 3-monooxygenase
-
Pseudomonas putida
HSP 3-monooxygenase
-
Pseudomonas putida
hspB
-
Pseudomonas putida

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
25
-
assay at Pseudomonas putida

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
5.49
-
NADH pH 8.0, 25°C, recombinant His-tagged enzyme Pseudomonas putida
7.74
-
4-(6-hydroxypyridin-3-yl)-4-oxobutanoate pH 8.0, 25°C, recombinant His-tagged enzyme Pseudomonas putida

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
8
-
assay at Pseudomonas putida

Cofactor

Cofactor Comment Organism Structure
FAD
-
Pseudomonas putida
NADH
-
Pseudomonas putida

General Information

General Information Comment Organism
evolution phylogenetic analysis reveals that HspB is the most closely related to two p-nitrophenol 4-monooxygenases, and the experimental results exhibit that p-nitrophenol is a substrate of HspB Pseudomonas putida
additional information free H2O2 does not catalyze the HspB enzyme reaction Pseudomonas putida
physiological function 6-hydroxy-3-succinoyl-pyridine (HSP) 3-monooxygenase (HspB) is a flavoprotein essential to the pyrrolidine pathway of nicotine degradation, it catalyzes pyridine-ring beta-hydroxylation, resulting in carbon-carbon cleavage and production of 2,5-dihydroxypyridine Pseudomonas putida