Heterologous overexpression and biochemical characterization of a nitroreductase from Gluconobacter oxydans 621H

Yang, Y.; Lin, J.; Wei, D.; Mol. Biotechnol. 58, 428-440 (2016)

Data extracted from this reference:

Inhibitors

Commentary

Organism

Structure

Cd2+

1 mM, no residual activity

Gluconobacter oxydans

glutathione

1 mM, 41% residual activity

Gluconobacter oxydans

menadione

1 mM, 61% residual activity

Gluconobacter oxydans

Sodium azide

1 mM, 52% residual activity

Gluconobacter oxydans

Triton X-100

1% w/v, 84% residual activity

Gluconobacter oxydans

Tween 20

1% w/v, 64% residual activity

Gluconobacter oxydans

ZnSO4

1 mM, no residual activity

Gluconobacter oxydans

KM Value [mM]

KM Value Maximum [mM]

Substrate

Commentary

Organism

Structure

0.016

-

(2E)-2-[(5-nitrofuran-2-yl)methylidene]hydrazine-1-carboxamide

pH 8.0, 37°C

Gluconobacter oxydans

0.054

-

5-(aziridin-1-yl)-2,4-dinitrobenzamide

pH 8.0, 37°C

Gluconobacter oxydans

0.128

-

1,3-dinitrobenzene

pH 8.0, 37°C

Gluconobacter oxydans

0.153

-

NADPH

pH 8.0, 37°C

Gluconobacter oxydans

0.264

-

3-nitrotoluene

pH 8.0, 37°C

Gluconobacter oxydans

0.282

-

4-nitrotoluene

pH 8.0, 37°C

Gluconobacter oxydans

0.309

-

4-chloronitrobenzene

pH 8.0, 37°C

Gluconobacter oxydans

0.468

-

2-chloronitrobenzene

pH 8.0, 37°C

Gluconobacter oxydans

0.545

-

nitrobenzene

pH 8.0, 37°C

Gluconobacter oxydans

Substrates

Commentary Substrates

Literature (Substrates)

Organism

Products

Commentary (Products)

Literature (Products)

Organism (Products)

Reversibility

Substrate Product ID

(2E)-2-[(5-nitrofuran-2-yl)methylidene]hydrazine-1-carboxamide + 2 NADPH + 2 H+

i.e. anti-infective furacilin

740989

Gluconobacter oxydans

(2E)-2-[[5-(hydroxyamino)furan-2-yl]methylidene]hydrazine-1-carboxamide + 2 NADP+

-

-

-

?

(2E)-2-[(5-nitrofuran-2-yl)methylidene]hydrazine-1-carboxamide + 2 NADPH + 2 H+

i.e. anti-infective furacilin

740989

Gluconobacter oxydans 621H

(2E)-2-[[5-(hydroxyamino)furan-2-yl]methylidene]hydrazine-1-carboxamide + 2 NADP+

-

-

-

?

1,3-dinitrobenzene + 2 NADPH + 2 H+

-

740989

Gluconobacter oxydans

? + 2 NADP+

-

-

-

?

1,3-dinitrobenzene + 2 NADPH + 2 H+

-

740989

Gluconobacter oxydans 621H

? + 2 NADP+

-

-

-

?

2-chloronitrobenzene + 2 NADPH + 2 H+

-

740989

Gluconobacter oxydans

2-chloro-N-hydroxyaniline + 2 NADP+ + H2O

-

-

-

?

2-chloronitrobenzene + 2 NADPH + 2 H+

-

740989

Gluconobacter oxydans 621H

2-chloro-N-hydroxyaniline + 2 NADP+ + H2O

-

-

-

?

3-nitrotoluene + 2 NADPH + 2 H+

-

740989

Gluconobacter oxydans

N-hydroxy-3-methylaniline + 2 NADP+ + H2O

-

-

-

?

3-nitrotoluene + 2 NADPH + 2 H+

-

740989

Gluconobacter oxydans 621H

N-hydroxy-3-methylaniline + 2 NADP+ + H2O

-

-

-

?

4-chloronitrobenzene + 2 NADPH + 2 H+

-

740989

Gluconobacter oxydans

4-chloro-N-hydroxyaniline + 2 NADP+ + H2O

-

-

-

?

4-nitrotoluene + 2 NADPH + 2 H+

-

740989

Gluconobacter oxydans

N-hydroxy-4-methylaniline + 2 NADP+ + H2O

-

-

-

?

5-(aziridin-1-yl)-2,4-dinitrobenzamide + 2 NADPH + 2 H+

-

740989

Gluconobacter oxydans

? + 2 NADP+

enzyme activates the dinitrobenzamide cancer prodrug CB1954, i.e. 5-(aziridin-1-yl)-2,4-dinitrobenzamide

-

-

?

nitrobenzene + 2 NADPH + 2 H+

-

740989

Gluconobacter oxydans

N-hydroxyaniline + 2 NADP+ + H2O

-

-

-

?

nitrobenzene + 2 NADPH + 2 H+

-

740989

Gluconobacter oxydans 621H

N-hydroxyaniline + 2 NADP+ + H2O

-

-

-

?

Turnover Number Minimum [1/s]

Turnover Number Maximum [1/s]

Substrate

Commentary

Organism

Structure

0.1

-

(2E)-2-[(5-nitrofuran-2-yl)methylidene]hydrazine-1-carboxamide

pH 8.0, 37°C

Gluconobacter oxydans

0.12

-

4-chloronitrobenzene

pH 8.0, 37°C

Gluconobacter oxydans

0.13

-

4-nitrotoluene

pH 8.0, 37°C

Gluconobacter oxydans

0.15

-

2-chloronitrobenzene

pH 8.0, 37°C

Gluconobacter oxydans

0.18

-

3-nitrotoluene

pH 8.0, 37°C

Gluconobacter oxydans

0.18

-

nitrobenzene

pH 8.0, 37°C

Gluconobacter oxydans

0.24

-

1,3-dinitrobenzene

pH 8.0, 37°C

Gluconobacter oxydans

1.05

-

NADPH

pH 8.0, 37°C

Gluconobacter oxydans

1.06

-

5-(aziridin-1-yl)-2,4-dinitrobenzamide

pH 8.0, 37°C

Gluconobacter oxydans

Cofactor

Commentary

Organism

Structure

NADPH

-

Gluconobacter oxydans

Cofactor

Commentary

Organism

Structure

NADPH

-

Gluconobacter oxydans

Inhibitors

Commentary

Organism

Structure

Cd2+

1 mM, no residual activity

Gluconobacter oxydans

glutathione

1 mM, 41% residual activity

Gluconobacter oxydans

menadione

1 mM, 61% residual activity

Gluconobacter oxydans

Sodium azide

1 mM, 52% residual activity

Gluconobacter oxydans

Triton X-100

1% w/v, 84% residual activity

Gluconobacter oxydans

Tween 20

1% w/v, 64% residual activity

Gluconobacter oxydans

ZnSO4

1 mM, no residual activity

Gluconobacter oxydans

KM Value [mM]

KM Value Maximum [mM]

Substrate

Commentary

Organism

Structure

0.016

-

(2E)-2-[(5-nitrofuran-2-yl)methylidene]hydrazine-1-carboxamide

pH 8.0, 37°C

Gluconobacter oxydans

0.054

-

5-(aziridin-1-yl)-2,4-dinitrobenzamide

pH 8.0, 37°C

Gluconobacter oxydans

0.128

-

1,3-dinitrobenzene

pH 8.0, 37°C

Gluconobacter oxydans

0.153

-

NADPH

pH 8.0, 37°C

Gluconobacter oxydans

0.264

-

3-nitrotoluene

pH 8.0, 37°C

Gluconobacter oxydans

0.282

-

4-nitrotoluene

pH 8.0, 37°C

Gluconobacter oxydans

0.309

-

4-chloronitrobenzene

pH 8.0, 37°C

Gluconobacter oxydans

0.468

-

2-chloronitrobenzene

pH 8.0, 37°C

Gluconobacter oxydans

0.545

-

nitrobenzene

pH 8.0, 37°C

Gluconobacter oxydans

Substrates

Commentary Substrates

Literature (Substrates)

Organism

Products

Commentary (Products)

Literature (Products)

Organism (Products)

Reversibility

ID

(2E)-2-[(5-nitrofuran-2-yl)methylidene]hydrazine-1-carboxamide + 2 NADPH + 2 H+

i.e. anti-infective furacilin

740989

Gluconobacter oxydans

(2E)-2-[[5-(hydroxyamino)furan-2-yl]methylidene]hydrazine-1-carboxamide + 2 NADP+

-

-

-

?

(2E)-2-[(5-nitrofuran-2-yl)methylidene]hydrazine-1-carboxamide + 2 NADPH + 2 H+

i.e. anti-infective furacilin

740989

Gluconobacter oxydans 621H

(2E)-2-[[5-(hydroxyamino)furan-2-yl]methylidene]hydrazine-1-carboxamide + 2 NADP+

-

-

-

?

1,3-dinitrobenzene + 2 NADPH + 2 H+

-

740989

Gluconobacter oxydans

? + 2 NADP+

-

-

-

?

1,3-dinitrobenzene + 2 NADPH + 2 H+

-

740989

Gluconobacter oxydans 621H

? + 2 NADP+

-

-

-

?

2-chloronitrobenzene + 2 NADPH + 2 H+

-

740989

Gluconobacter oxydans

2-chloro-N-hydroxyaniline + 2 NADP+ + H2O

-

-

-

?

2-chloronitrobenzene + 2 NADPH + 2 H+

-

740989

Gluconobacter oxydans 621H

2-chloro-N-hydroxyaniline + 2 NADP+ + H2O

-

-

-

?

3-nitrotoluene + 2 NADPH + 2 H+

-

740989

Gluconobacter oxydans

N-hydroxy-3-methylaniline + 2 NADP+ + H2O

-

-

-

?

3-nitrotoluene + 2 NADPH + 2 H+

-

740989

Gluconobacter oxydans 621H

N-hydroxy-3-methylaniline + 2 NADP+ + H2O

-

-

-

?

4-chloronitrobenzene + 2 NADPH + 2 H+

-

740989

Gluconobacter oxydans

4-chloro-N-hydroxyaniline + 2 NADP+ + H2O

-

-

-

?

4-nitrotoluene + 2 NADPH + 2 H+

-

740989

Gluconobacter oxydans

N-hydroxy-4-methylaniline + 2 NADP+ + H2O

-

-

-

?

5-(aziridin-1-yl)-2,4-dinitrobenzamide + 2 NADPH + 2 H+

-

740989

Gluconobacter oxydans

? + 2 NADP+

enzyme activates the dinitrobenzamide cancer prodrug CB1954, i.e. 5-(aziridin-1-yl)-2,4-dinitrobenzamide

-

-

?

nitrobenzene + 2 NADPH + 2 H+

-

740989

Gluconobacter oxydans

N-hydroxyaniline + 2 NADP+ + H2O

-

-

-

?

nitrobenzene + 2 NADPH + 2 H+

-

740989

Gluconobacter oxydans 621H

N-hydroxyaniline + 2 NADP+ + H2O

-

-

-

?

Turnover Number Minimum [1/s]

Turnover Number Maximum [1/s]

Substrate

Commentary

Organism

Structure

0.1

-

(2E)-2-[(5-nitrofuran-2-yl)methylidene]hydrazine-1-carboxamide

pH 8.0, 37°C

Gluconobacter oxydans

0.12

-

4-chloronitrobenzene

pH 8.0, 37°C

Gluconobacter oxydans

0.13

-

4-nitrotoluene

pH 8.0, 37°C

Gluconobacter oxydans

0.15

-

2-chloronitrobenzene

pH 8.0, 37°C

Gluconobacter oxydans

0.18

-

3-nitrotoluene

pH 8.0, 37°C

Gluconobacter oxydans

0.18

-

nitrobenzene

pH 8.0, 37°C

Gluconobacter oxydans

0.24

-

1,3-dinitrobenzene

pH 8.0, 37°C

Gluconobacter oxydans

1.05

-

NADPH

pH 8.0, 37°C

Gluconobacter oxydans

1.06

-

5-(aziridin-1-yl)-2,4-dinitrobenzamide

pH 8.0, 37°C

Gluconobacter oxydans

kcat/KM Value [1/mMs^-1]

kcat/KM Value Maximum [1/mMs^-1]

Substrate

Commentary

Organism

Structure

0.32

-

2-chloronitrobenzene

pH 8.0, 37°C

Gluconobacter oxydans

0.33

-

nitrobenzene

pH 8.0, 37°C

Gluconobacter oxydans

0.39

-

4-chloronitrobenzene

pH 8.0, 37°C

Gluconobacter oxydans

0.46

-

4-nitrotoluene

pH 8.0, 37°C

Gluconobacter oxydans

0.7

-

3-nitrotoluene

pH 8.0, 37°C

Gluconobacter oxydans

1.9

-

1,3-dinitrobenzene

pH 8.0, 37°C

Gluconobacter oxydans

6.9

-

NADPH

pH 8.0, 37°C

Gluconobacter oxydans

20

-

5-(aziridin-1-yl)-2,4-dinitrobenzamide

pH 8.0, 37°C

Gluconobacter oxydans

60

-

(2E)-2-[(5-nitrofuran-2-yl)methylidene]hydrazine-1-carboxamide

pH 8.0, 37°C

Gluconobacter oxydans

KCat/KM Value [1/mMs^-1]

KCat/KM Value Maximum [1/mMs^-1]

Substrate

Commentary

Organism

Structure

0.32

-

2-chloronitrobenzene

pH 8.0, 37°C

Gluconobacter oxydans

0.33

-

nitrobenzene

pH 8.0, 37°C

Gluconobacter oxydans

0.39

-

4-chloronitrobenzene

pH 8.0, 37°C

Gluconobacter oxydans

0.46

-

4-nitrotoluene

pH 8.0, 37°C

Gluconobacter oxydans

0.7

-

3-nitrotoluene

pH 8.0, 37°C

Gluconobacter oxydans

1.9

-

1,3-dinitrobenzene

pH 8.0, 37°C

Gluconobacter oxydans

6.9

-

NADPH

pH 8.0, 37°C

Gluconobacter oxydans

20

-

5-(aziridin-1-yl)-2,4-dinitrobenzamide

pH 8.0, 37°C

Gluconobacter oxydans

60

-

(2E)-2-[(5-nitrofuran-2-yl)methylidene]hydrazine-1-carboxamide

pH 8.0, 37°C

Gluconobacter oxydans

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

Synonyms

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)

740989

Yang

Heterologous overexpression an ...

Gluconobacter oxydans, Gluconobacter oxydans 621H

Mol. Biotechnol.

58

428-440

2016

-

-

-

-

-

-

7

9

-

-

-

-

-

2

-

-

-

-

-

-

1

-

13

-

-

-

-

-

9

-

-

-

1

-

-

-

-

-

-

1

-

-

-

-

7

-

9

-

-

-

-

-

-

-

-

-

-

1

-

13

-

-

-

-

9

-

-

-

-

-

-

-

-

9

9

740192

Li

In vivo imaging and detection ...

Danio rerio

Biosens. Bioelectron.

63

112-116

2015

-

1

1

-

-

-

-

-

-

-

-

-

-

1

-

-

-

-

-

1

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

1

1

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

1

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

740325

Yuan

-

Isolation and characterization ...

Pseudomonas sp., Pseudomonas sp. N1

Chin. J. Appl. Environ. Biol.

19

1031-1034

2013

-

1

1

-

-

-

-

-

-

-

-

-

-

2

-

-

-

-

-

-

-

-

2

-

-

-

-

-

-

-

-

-

-

-

-

-

-

1

1

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

2

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

741380

Isayev

In silico structure-function a ...

Enterobacter cloacae

Proteins

80

2728-2741

2012

-

-

-

1

-

-

-

-

-

-

-

-

-

1

-

-

-

-

-

-

-

-

-

-

1

-

-

-

-

-

-

-

1

-

-

-

-

-

-

1

1

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

741093

Yanto

Nitroreductase from Salmonella ...

Salmonella enterica subsp. enterica serovar Typhimurium, Salmonella enterica subsp. enterica serovar Typhimurium LT2

Org. Biomol. Chem.

8

1826-1832

2010

-

-

-

-

-

-

-

-

-

-

-

-

-

2

-

-

-

-

-

-

2

-

6

-

1

-

-

3

-

1

1

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

2

-

6

-

-

-

3

-

1

1

-

-

-

-

-

-

-

-

677659

Wu

Novel partial reductive pathwa ...

Comamonas sp., Comamonas sp. Ctestosteroni

Appl. Environ. Microbiol.

72

1759-1765

2006

-

-

1

-

-

-

-

-

-

-

-

-

-

2

-

-

-

-

-

-

-

-

2

-

1

-

-

-

-

-

-

-

1

-

-

-

-

-

1

1

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

2

-

-

-

-

-

-

-

-

-

-

1

1

-

-

-

677735

Xiao

Characterization of genes invo ...

Pseudomonas putida, Pseudomonas putida ZWL73

Appl. Microbiol. Biotechnol.

73

166-171

2006

-

-

1

-

-

-

-

-

-

-

-

-

-

7

-

-

-

-

-

-

-

-

14

-

1

-

-

-

-

-

-

-

2

-

-

-

-

-

1

2

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

14

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

740133

Berne

Application of a microfluidic ...

Pseudomonas pseudoalcaligenes, Pseudomonas pseudoalcaligenes JS45

Biomacromolecules

7

2631-2636

2006

-

1

1

-

-

-

-

6

-

-

-

-

-

4

-

-

1

-

-

-

-

-

6

-

1

-

-

-

-

-

-

-

-

-

-

-

-

1

1

-

-

-

-

-

-

-

6

-

-

-

-

-

-

-

1

-

-

-

-

6

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

740929

Ho

-

Characterization of nitroreduc ...

Pseudomonas sp. HK-6

Korean J. Microbiol. Biotechnol.

32

230-237

2004

-

-

-

-

-

-

5

-

-

-

1

-

-

1

-

-

1

-

-

-

1

-

2

1

-

1

-

-

-

1

-

-

-

-

-

-

-

-

-

-

-

-

-

-

5

-

-

-

-

1

-

-

-

-

1

-

-

1

-

2

1

1

-

-

-

1

-

-

-

-

-

-

-

-

-

740596

Somerville

Purification and characterizat ...

Pseudomonas pseudoalcaligenes, Pseudomonas pseudoalcaligenes JS45

J. Bacteriol.

177

3837-3842

1995

1

-

-

-

-

-

11

3

-

-

2

4

-

4

-

-

1

-

-

-

1

-

8

1

-

-

-

-

-

-

-

-

3

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1

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3

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11

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4

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8

1

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739974

Rieble

Aromatic nitroreductase from t ...

Phanerochaete chrysosporium

Biochem. Biophys. Res. Commun.

205

298-304

1994

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