Azoreductase (gene AzoRed2) from Streptomyces sp. S27

Enzyme Description

Species

Extremophile

EC Number

1.7.1.6 ↗

Sequence

Length: 233 amino acids

VPRLLHLDSSARTDSFTRRLGAAFVRAWRLNGGGEVTHRDLAIDPPPPISEGWTSLCDTLLREGITEAATDHYAQAVRTEAERRAWAVTEPLLAELLAADVVLIGAPMYNFGVPAALKAWIDQVTFPKMVLAPRRFVVLSARGGAYGPGTPREPFDHQGRYLLDFFHGHYGVEDAEVISAELVNSLSDPGLAGRLPQHLDSAAAALARAVELGTELATVPATTSADRTLAKEG

Hao Dong et al. (2019) — Biochemical characterization of a novel azoreductase from Streptomyces sp.: Application in eco-friendly decolorization of azo dye wastewater

International Journal of Biological Macromolecules · doi:10.1016/j.ijbiomac.2019.08.196 ↗ · Stability - Incubation

22 measurements ▼

Database ID

UniParc: UPI0012A9B19E ↗

Sequence Annotation

Explicit - Provided GenBank Accession Number

Protein Source

Recombinant, host bacterium Escherichia coli BL21 (DE3)

Experimental Data (22 measurements)

22 measurements — page 2 of 2

Property	Assay	Solvent	Solvent Volume	Aqueous Reference	Measured Value	Units	Solution	pH	Temperature	Substrate(s)	Product(s)	Cofactor(s)	Shaking	Comments
Stability - Incubation	Activity after incubation (1 hour at 30°C), measured by absorbance spectrophotometry (decrease in methyl red absorbance measurement, 430 nm) in aqueous phase	Ethanol	10%	100.0	91.41	%	Incubation: ?, Assay: 25 mM Tris-HCl buffer	Incubation: ?, Assay: 7.4	Incubation: 30°C, Assay: 30°C	25 µM Methyl red	N,N-dimethyl-p-phenylenediamine , p-aminobenzoic acid	250 µM NADH, 5 µM FMN	Incubation: 80 rpm	Non-incubated control (in %), assay in aqueous phase \| Clear assay in aqueous phase, still 5% organic solvent in assay solution for hydrophilic solvents
Stability - Incubation	Activity after incubation (1 hour at 30°C), measured by absorbance spectrophotometry (decrease in methyl red absorbance measurement, 430 nm) in aqueous phase	Methanol	10%	100.0	80.98	%	Incubation: ?, Assay: 25 mM Tris-HCl buffer	Incubation: ?, Assay: 7.4	Incubation: 30°C, Assay: 30°C	25 µM Methyl red	N,N-dimethyl-p-phenylenediamine , p-aminobenzoic acid	250 µM NADH, 5 µM FMN	Incubation: 80 rpm	Non-incubated control (in %), assay in aqueous phase \| Clear assay in aqueous phase, still 5% organic solvent in assay solution for hydrophilic solvents

Stability - Incubation

Activity after incubation (1 hour at 30°C), measured by absorbance spectrophotometry (decrease in methyl red absorbance measurement, 430 nm) in aqueous phase

Dimethyl Sulfoxide (DMSO)

20%

100.0

Incubation: ?, Assay: 25 mM Tris-HCl buffer

Incubation: ?, Assay: 7.4

Incubation: 30°C, Assay: 30°C

25 µM Methyl red

N,N-dimethyl-p-phenylenediamine, p-aminobenzoic acid

250 µM NADH, 5 µM FMN

Incubation: 80 rpm

Non-incubated control (in %), assay in aqueous phase | Clear assay in aqueous phase, still 5% organic solvent in assay solution for hydrophilic solvents

Stability - Incubation

Activity after incubation (1 hour at 30°C), measured by absorbance spectrophotometry (decrease in methyl red absorbance measurement, 430 nm) in aqueous phase

Isooctane

20%

100.0

91.99

Incubation: ?, Assay: 25 mM Tris-HCl buffer

Incubation: ?, Assay: 7.4

Incubation: 30°C, Assay: 30°C

25 µM Methyl red

N,N-dimethyl-p-phenylenediamine, p-aminobenzoic acid

250 µM NADH, 5 µM FMN

Incubation: 80 rpm

Non-incubated control (in %), assay in aqueous phase | Clear assay in aqueous phase, still 5% organic solvent in assay solution for hydrophilic solvents

Stability - Incubation

Activity after incubation (1 hour at 30°C), measured by absorbance spectrophotometry (decrease in methyl red absorbance measurement, 430 nm) in aqueous phase

n-Hexane

20%

100.0

92.33

Incubation: ?, Assay: 25 mM Tris-HCl buffer

Incubation: ?, Assay: 7.4

Incubation: 30°C, Assay: 30°C

25 µM Methyl red

N,N-dimethyl-p-phenylenediamine, p-aminobenzoic acid

250 µM NADH, 5 µM FMN

Incubation: 80 rpm

Non-incubated control (in %), assay in aqueous phase | Clear assay in aqueous phase, still 5% organic solvent in assay solution for hydrophilic solvents

Stability - Incubation

Activity after incubation (1 hour at 30°C), measured by absorbance spectrophotometry (decrease in methyl red absorbance measurement, 430 nm) in aqueous phase

Chloroform

20%

100.0

12.2

Incubation: ?, Assay: 25 mM Tris-HCl buffer

Incubation: ?, Assay: 7.4

Incubation: 30°C, Assay: 30°C

25 µM Methyl red

N,N-dimethyl-p-phenylenediamine, p-aminobenzoic acid

250 µM NADH, 5 µM FMN

Incubation: 80 rpm

Non-incubated control (in %), assay in aqueous phase | Clear assay in aqueous phase, still 5% organic solvent in assay solution for hydrophilic solvents

Stability - Incubation

Activity after incubation (1 hour at 30°C), measured by absorbance spectrophotometry (decrease in methyl red absorbance measurement, 430 nm) in aqueous phase

Isoamyl Alcohol

20%

100.0

9.76

Incubation: ?, Assay: 25 mM Tris-HCl buffer

Incubation: ?, Assay: 7.4

Incubation: 30°C, Assay: 30°C

25 µM Methyl red

N,N-dimethyl-p-phenylenediamine, p-aminobenzoic acid

250 µM NADH, 5 µM FMN

Incubation: 80 rpm

Non-incubated control (in %), assay in aqueous phase | Clear assay in aqueous phase, still 5% organic solvent in assay solution for hydrophilic solvents

Stability - Incubation

Activity after incubation (1 hour at 30°C), measured by absorbance spectrophotometry (decrease in methyl red absorbance measurement, 430 nm) in aqueous phase

1-Butanol

20%

100.0

6.27

Incubation: ?, Assay: 25 mM Tris-HCl buffer

Incubation: ?, Assay: 7.4

Incubation: 30°C, Assay: 30°C

25 µM Methyl red

N,N-dimethyl-p-phenylenediamine, p-aminobenzoic acid

250 µM NADH, 5 µM FMN

Incubation: 80 rpm

Non-incubated control (in %), assay in aqueous phase | Clear assay in aqueous phase, still 5% organic solvent in assay solution for hydrophilic solvents

Stability - Incubation

Activity after incubation (1 hour at 30°C), measured by absorbance spectrophotometry (decrease in methyl red absorbance measurement, 430 nm) in aqueous phase

1-Propanol

20%

100.0

23.73

Incubation: ?, Assay: 25 mM Tris-HCl buffer

Incubation: ?, Assay: 7.4

Incubation: 30°C, Assay: 30°C

25 µM Methyl red

N,N-dimethyl-p-phenylenediamine, p-aminobenzoic acid

250 µM NADH, 5 µM FMN

Incubation: 80 rpm

Non-incubated control (in %), assay in aqueous phase | Clear assay in aqueous phase, still 5% organic solvent in assay solution for hydrophilic solvents

Stability - Incubation

Activity after incubation (1 hour at 30°C), measured by absorbance spectrophotometry (decrease in methyl red absorbance measurement, 430 nm) in aqueous phase

Isopropanol

20%

100.0

109.32

Incubation: ?, Assay: 25 mM Tris-HCl buffer

Incubation: ?, Assay: 7.4

Incubation: 30°C, Assay: 30°C

25 µM Methyl red

N,N-dimethyl-p-phenylenediamine, p-aminobenzoic acid

250 µM NADH, 5 µM FMN

Incubation: 80 rpm

Non-incubated control (in %), assay in aqueous phase | Clear assay in aqueous phase, still 5% organic solvent in assay solution for hydrophilic solvents

Stability - Incubation

Activity after incubation (1 hour at 30°C), measured by absorbance spectrophotometry (decrease in methyl red absorbance measurement, 430 nm) in aqueous phase

Acetonitrile

20%

100.0

17.8

Incubation: ?, Assay: 25 mM Tris-HCl buffer

Incubation: ?, Assay: 7.4

Incubation: 30°C, Assay: 30°C

25 µM Methyl red

N,N-dimethyl-p-phenylenediamine, p-aminobenzoic acid

250 µM NADH, 5 µM FMN

Incubation: 80 rpm

Non-incubated control (in %), assay in aqueous phase | Clear assay in aqueous phase, still 5% organic solvent in assay solution for hydrophilic solvents

Stability - Incubation

Activity after incubation (1 hour at 30°C), measured by absorbance spectrophotometry (decrease in methyl red absorbance measurement, 430 nm) in aqueous phase

Ethanol

20%

100.0

105.93

Incubation: ?, Assay: 25 mM Tris-HCl buffer

Incubation: ?, Assay: 7.4

Incubation: 30°C, Assay: 30°C

25 µM Methyl red

N,N-dimethyl-p-phenylenediamine, p-aminobenzoic acid

250 µM NADH, 5 µM FMN

Incubation: 80 rpm

Non-incubated control (in %), assay in aqueous phase | Clear assay in aqueous phase, still 5% organic solvent in assay solution for hydrophilic solvents

Stability - Incubation

Activity after incubation (1 hour at 30°C), measured by absorbance spectrophotometry (decrease in methyl red absorbance measurement, 430 nm) in aqueous phase

Methanol

20%

100.0

Incubation: ?, Assay: 25 mM Tris-HCl buffer

Incubation: ?, Assay: 7.4

Incubation: 30°C, Assay: 30°C

25 µM Methyl red

N,N-dimethyl-p-phenylenediamine, p-aminobenzoic acid

250 µM NADH, 5 µM FMN

Incubation: 80 rpm

Non-incubated control (in %), assay in aqueous phase | Clear assay in aqueous phase, still 5% organic solvent in assay solution for hydrophilic solvents

Stability - Incubation

Activity after incubation (1 hour at 30°C), measured by absorbance spectrophotometry (decrease in methyl red absorbance measurement, 430 nm) in aqueous phase

Dimethyl Sulfoxide (DMSO)

10%

100.0

85.28

Incubation: ?, Assay: 25 mM Tris-HCl buffer

Incubation: ?, Assay: 7.4

Incubation: 30°C, Assay: 30°C

25 µM Methyl red

N,N-dimethyl-p-phenylenediamine, p-aminobenzoic acid

250 µM NADH, 5 µM FMN

Incubation: 80 rpm

Non-incubated control (in %), assay in aqueous phase | Clear assay in aqueous phase, still 5% organic solvent in assay solution for hydrophilic solvents

Stability - Incubation

Activity after incubation (1 hour at 30°C), measured by absorbance spectrophotometry (decrease in methyl red absorbance measurement, 430 nm) in aqueous phase

Isooctane

10%

100.0

69.03

Incubation: ?, Assay: 25 mM Tris-HCl buffer

Incubation: ?, Assay: 7.4

Incubation: 30°C, Assay: 30°C

25 µM Methyl red

N,N-dimethyl-p-phenylenediamine, p-aminobenzoic acid

250 µM NADH, 5 µM FMN

Incubation: 80 rpm

Non-incubated control (in %), assay in aqueous phase | Clear assay in aqueous phase, still 5% organic solvent in assay solution for hydrophilic solvents

Stability - Incubation

Activity after incubation (1 hour at 30°C), measured by absorbance spectrophotometry (decrease in methyl red absorbance measurement, 430 nm) in aqueous phase

n-Hexane

10%

100.0

103.73

Incubation: ?, Assay: 25 mM Tris-HCl buffer

Incubation: ?, Assay: 7.4

Incubation: 30°C, Assay: 30°C

25 µM Methyl red

N,N-dimethyl-p-phenylenediamine, p-aminobenzoic acid

250 µM NADH, 5 µM FMN

Incubation: 80 rpm

Non-incubated control (in %), assay in aqueous phase | Clear assay in aqueous phase, still 5% organic solvent in assay solution for hydrophilic solvents

Stability - Incubation

Activity after incubation (1 hour at 30°C), measured by absorbance spectrophotometry (decrease in methyl red absorbance measurement, 430 nm) in aqueous phase

Chloroform

10%

100.0

10.82

Incubation: ?, Assay: 25 mM Tris-HCl buffer

Incubation: ?, Assay: 7.4

Incubation: 30°C, Assay: 30°C

25 µM Methyl red

N,N-dimethyl-p-phenylenediamine, p-aminobenzoic acid

250 µM NADH, 5 µM FMN

Incubation: 80 rpm

Non-incubated control (in %), assay in aqueous phase | Clear assay in aqueous phase, still 5% organic solvent in assay solution for hydrophilic solvents

Stability - Incubation

Activity after incubation (1 hour at 30°C), measured by absorbance spectrophotometry (decrease in methyl red absorbance measurement, 430 nm) in aqueous phase

Isoamyl Alcohol

10%

100.0

13.06

Incubation: ?, Assay: 25 mM Tris-HCl buffer

Incubation: ?, Assay: 7.4

Incubation: 30°C, Assay: 30°C

25 µM Methyl red

N,N-dimethyl-p-phenylenediamine, p-aminobenzoic acid

250 µM NADH, 5 µM FMN

Incubation: 80 rpm

Non-incubated control (in %), assay in aqueous phase | Clear assay in aqueous phase, still 5% organic solvent in assay solution for hydrophilic solvents

Stability - Incubation

Activity after incubation (1 hour at 30°C), measured by absorbance spectrophotometry (decrease in methyl red absorbance measurement, 430 nm) in aqueous phase

1-Butanol

10%

100.0

13.06

Incubation: ?, Assay: 25 mM Tris-HCl buffer

Incubation: ?, Assay: 7.4

Incubation: 30°C, Assay: 30°C

25 µM Methyl red

N,N-dimethyl-p-phenylenediamine, p-aminobenzoic acid

250 µM NADH, 5 µM FMN

Incubation: 80 rpm

Non-incubated control (in %), assay in aqueous phase | Clear assay in aqueous phase, still 5% organic solvent in assay solution for hydrophilic solvents

Stability - Incubation

Activity after incubation (1 hour at 30°C), measured by absorbance spectrophotometry (decrease in methyl red absorbance measurement, 430 nm) in aqueous phase

1-Propanol

10%

100.0

84.05

Incubation: ?, Assay: 25 mM Tris-HCl buffer

Incubation: ?, Assay: 7.4

Incubation: 30°C, Assay: 30°C

25 µM Methyl red

N,N-dimethyl-p-phenylenediamine, p-aminobenzoic acid

250 µM NADH, 5 µM FMN

Incubation: 80 rpm

Non-incubated control (in %), assay in aqueous phase | Clear assay in aqueous phase, still 5% organic solvent in assay solution for hydrophilic solvents

Stability - Incubation

Activity after incubation (1 hour at 30°C), measured by absorbance spectrophotometry (decrease in methyl red absorbance measurement, 430 nm) in aqueous phase

Isopropanol

10%

100.0

87.73

Incubation: ?, Assay: 25 mM Tris-HCl buffer

Incubation: ?, Assay: 7.4

Incubation: 30°C, Assay: 30°C

25 µM Methyl red

N,N-dimethyl-p-phenylenediamine, p-aminobenzoic acid

250 µM NADH, 5 µM FMN

Incubation: 80 rpm

Non-incubated control (in %), assay in aqueous phase | Clear assay in aqueous phase, still 5% organic solvent in assay solution for hydrophilic solvents

Stability - Incubation

Activity after incubation (1 hour at 30°C), measured by absorbance spectrophotometry (decrease in methyl red absorbance measurement, 430 nm) in aqueous phase

Acetonitrile

10%

100.0

48.47

Incubation: ?, Assay: 25 mM Tris-HCl buffer

Incubation: ?, Assay: 7.4

Incubation: 30°C, Assay: 30°C

25 µM Methyl red

N,N-dimethyl-p-phenylenediamine, p-aminobenzoic acid

250 µM NADH, 5 µM FMN

Incubation: 80 rpm

Non-incubated control (in %), assay in aqueous phase | Clear assay in aqueous phase, still 5% organic solvent in assay solution for hydrophilic solvents

Stability - Incubation

Activity after incubation (1 hour at 30°C), measured by absorbance spectrophotometry (decrease in methyl red absorbance measurement, 430 nm) in aqueous phase

Ethanol

10%

100.0

91.41

Incubation: ?, Assay: 25 mM Tris-HCl buffer

Incubation: ?, Assay: 7.4

Incubation: 30°C, Assay: 30°C

25 µM Methyl red

N,N-dimethyl-p-phenylenediamine, p-aminobenzoic acid

250 µM NADH, 5 µM FMN

Incubation: 80 rpm

Non-incubated control (in %), assay in aqueous phase | Clear assay in aqueous phase, still 5% organic solvent in assay solution for hydrophilic solvents

Stability - Incubation

Activity after incubation (1 hour at 30°C), measured by absorbance spectrophotometry (decrease in methyl red absorbance measurement, 430 nm) in aqueous phase

Methanol

10%

100.0

80.98

Incubation: ?, Assay: 25 mM Tris-HCl buffer

Incubation: ?, Assay: 7.4

Incubation: 30°C, Assay: 30°C

25 µM Methyl red

N,N-dimethyl-p-phenylenediamine, p-aminobenzoic acid

250 µM NADH, 5 µM FMN

Incubation: 80 rpm

Non-incubated control (in %), assay in aqueous phase | Clear assay in aqueous phase, still 5% organic solvent in assay solution for hydrophilic solvents

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Visualization : Stability — Incubation

One bar per measurement. Colour = solvent, shade = solvent volume.

Structure

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Azoreductase (gene AzoRed2) from Streptomyces sp. S27

Enzyme Description

Sequence

Experimental Data (22 measurements)

Visualization : Stability — Incubation

Structure

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