BRENDA - Reference to 1.14.13.163; Id = 745310

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

Pseudomonas putida

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

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

-

?

Pseudomonas putida

-

Pseudomonas putida S16

-

recombinant His6-tagged enzyme from Escherichia coli strain BL21(DE3) by nickel affinity chromatography

Pseudomonas putida

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

-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

4-(6-hydroxypyridin-3-yl)-4-oxobutanoate + 2 NADH + 2 H+ + O2

-

745310

Pseudomonas putida

2,5-dihydroxypyridine + succinate semialdehyde + 2 NAD+ + H2O

-

?

4-(6-hydroxypyridin-3-yl)-4-oxobutanoate + 2 NADH + 2 H+ + O2

-

745310

Pseudomonas putida S16

2,5-dihydroxypyridine + succinate semialdehyde + 2 NAD+ + H2O

-

?

additional information

mass spectrometric analysis of substrates and products, with recombinant enzyme

745310

Pseudomonas putida

?

-

additional information

mass spectrometric analysis of substrates and products, with recombinant enzyme

745310

Pseudomonas putida S16

?

-

25

-

assay at

Pseudomonas putida

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

8

-

assay at

Pseudomonas putida

FAD

-

Pseudomonas putida

NADH

-

Pseudomonas putida

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

Pseudomonas putida

FAD

-

Pseudomonas putida

NADH

-

Pseudomonas putida

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

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

-

?

recombinant His6-tagged enzyme from Escherichia coli strain BL21(DE3) by nickel affinity chromatography

Pseudomonas putida

-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

4-(6-hydroxypyridin-3-yl)-4-oxobutanoate + 2 NADH + 2 H+ + O2

-

745310

Pseudomonas putida

2,5-dihydroxypyridine + succinate semialdehyde + 2 NAD+ + H2O

-

?

4-(6-hydroxypyridin-3-yl)-4-oxobutanoate + 2 NADH + 2 H+ + O2

-

745310

Pseudomonas putida S16

2,5-dihydroxypyridine + succinate semialdehyde + 2 NAD+ + H2O

-

?

additional information

mass spectrometric analysis of substrates and products, with recombinant enzyme

745310

Pseudomonas putida

?

-

additional information

mass spectrometric analysis of substrates and products, with recombinant enzyme

745310

Pseudomonas putida S16

?

-

25

-

assay at

Pseudomonas putida

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

8

-

assay at

Pseudomonas putida

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

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

744640

Wei

Characterization of a novel n ...

Pseudomonas sp. ZZ-5

Catalysts

7

272-281

2017

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1

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18

2

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1

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1

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1

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1

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1

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18

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2

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1

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1

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1

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3

1

2

1

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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

-

741691

Yu

Molecular mechanism of nicoti ...

Ochrobactrum sp.

Appl. Environ. Microbiol.

81

272-281

2015

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1

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3

2

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1

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1

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3

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745310

Yu

Mechanism of the 6-hydroxy-3- ...

Pseudomonas putida, Pseudomonas putida S16

J. Biol. Chem.

289

29158-29170

2014

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1

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3

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2

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4

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1

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1

4

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1

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1

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1

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1

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1

4

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1

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1

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3

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746248

Li

6-hydroxy-3-succinoylpyridine ...

Agrobacterium tumefaciens, Agrobacterium tumefaciens S33

PLoS ONE

9

e103324

2014

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1

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1

2

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7

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1

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2

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6

1

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1

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6

1

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1

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2

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718677

Wang

Identification of nicotine bio ...

Agrobacterium tumefaciens, Agrobacterium tumefaciens S33

Appl. Microbiol. Biotechnol.

95

1567-1578

2012

-

9

-

2

-

2

-

1

-

719982

Tang

A novel NADH-dependent and FAD ...

Pseudomonas putida, Pseudomonas putida S16

J. Biol. Chem.

286

39179-39187

2011

1

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1

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2

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2

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9

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690527

Tang

A novel gene, encoding 6-hydro ...

Pseudomonas putida, Pseudomonas putida S16

Appl. Environ. Microbiol.

74

1567-1574

2008

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1

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1

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5

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2

1

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1

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1

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1

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1

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Mechanism of the 6-hydroxy-3-succinoyl-pyridine 3-monooxygenase flavoprotein from Pseudomonas putida S16

Data extracted from this reference: