Haloperoxidase BPO-A1 from Streptomyces aureofaciens

Enzyme Description

Extremophile
No
EC Number

Sequence

Length: 275 amino acids
MPICTTRDGVEIFYKDWGQGRPVVFIHGWPLNGDAWQDQLKAVVDAGYRGIAHDRRGHGHSTPVWDGYDFDTFADDLNDLLTDLDLRDVTLVAHSMGGGELARYVGRHGTGRLRSAVLLSAIPPVMIKSDKNPDGVPDEVFDALKNGVLTERSQFWKDTAEGFFSANRPGNKVTQGNKDAFWYMAMAQTIEGGVRCVDAFGYTDFTEDLKKFDIPTLVVHGDDDQVVPIDATGRKSAQIIPNAELKVYEGSSHGIAMVPGDKEKFNRDLLEFLNK
Ryosuke Yamada et al. (2014) β€” Improvement of the stability and activity of the BPO-A1 haloperoxidase from Streptomyces aureofaciens by directed evolution
Journal of Biotechnology  Β· doi:10.1016/j.jbiotec.2014.10.030 β†—  Β· Stability - Incubation
31 measurements
Database ID
UniProt: P33912 β†—
Sequence Annotation
Explicit - Provided
Protein Source
Recombinant, host bacterium Escherichia coli Rosetta2 (DE3)

Experimental Data (31 measurements)

31 measurements β€” page 2 of 2
Mutation Property Assay Solvent Solvent Volume Aqueous ReferenceWT Reference Measured Value Units Extraction method Data type Solution pH Temperature Substrate(s) Product(s) Cofactor(s) Shaking Comments
R114H/N146H Stability - Incubation Activity after incubation (48 hours at 30Β°C), measured by absorbance spectrophotometry (colorimetric assay, chlorodimedone and HOBr reaction product absorbance measurement, 290 nm) in aqueous phase Methanol 40 % v/v 100.0 89.0 104.0 % Digital Continuous Incubation: ?, Assay: 1 M sodium acetate buffer, 10 mM NaN3, 50 Β΅M Chlorodimedone Incubation: ?, Assay: 5.5 Incubation: 30Β°C, Assay: 25Β°C 10 mM Hβ‚‚Oβ‚‚ (Hydrogen Peroxide), 100 mM Sodium bromide H2O , HOBr β€” β€” Water-incubated control (in %), assay in aqueous phase | Uncertainty about whether assay was in the presence of organic solvent or in aqueous phase
G106S/V148I Stability - Incubation Activity after incubation (1 hour at 30Β°C), measured by absorbance spectrophotometry (colorimetric assay, chlorodimedone and HOBr reaction product absorbance measurement, 290 nm) in aqueous phase 1-Butanol 50 % v/v 96.0 6.0 11.0 % Digital Continuous Incubation: ?, Assay: 1 M sodium acetate buffer, 10 mM NaN3, 50 Β΅M Chlorodimedone Incubation: ?, Assay: 5.5 Incubation: 30Β°C, Assay: 25Β°C 10 mM Hβ‚‚Oβ‚‚ (Hydrogen Peroxide), 100 mM Sodium bromide H2O , HOBr β€” β€” Water-incubated control (in %), assay in aqueous phase | Uncertainty about whether assay was in the presence of organic solvent or in aqueous phase
R114H/N146H Stability - Incubation Activity after incubation (1 hour at 30Β°C), measured by absorbance spectrophotometry (colorimetric assay, chlorodimedone and HOBr reaction product absorbance measurement, 290 nm) in aqueous phase 1-Butanol 50 % v/v 100.0 6.0 18.0 % Digital Continuous Incubation: ?, Assay: 1 M sodium acetate buffer, 10 mM NaN3, 50 Β΅M Chlorodimedone Incubation: ?, Assay: 5.5 Incubation: 30Β°C, Assay: 25Β°C 10 mM Hβ‚‚Oβ‚‚ (Hydrogen Peroxide), 100 mM Sodium bromide H2O , HOBr β€” β€” Water-incubated control (in %), assay in aqueous phase | Uncertainty about whether assay was in the presence of organic solvent or in aqueous phase
G106S/V148I Stability - Incubation Activity after incubation (1 hour at 30Β°C), measured by absorbance spectrophotometry (colorimetric assay, chlorodimedone and HOBr reaction product absorbance measurement, 290 nm) in aqueous phase 1-Butanol 40 % v/v 96.0 26.0 15.0 % Digital Continuous Incubation: ?, Assay: 1 M sodium acetate buffer, 10 mM NaN3, 50 Β΅M Chlorodimedone Incubation: ?, Assay: 5.5 Incubation: 30Β°C, Assay: 25Β°C 10 mM Hβ‚‚Oβ‚‚ (Hydrogen Peroxide), 100 mM Sodium bromide H2O , HOBr β€” β€” Water-incubated control (in %), assay in aqueous phase | Uncertainty about whether assay was in the presence of organic solvent or in aqueous phase
R114H/N146H Stability - Incubation Activity after incubation (1 hour at 30Β°C), measured by absorbance spectrophotometry (colorimetric assay, chlorodimedone and HOBr reaction product absorbance measurement, 290 nm) in aqueous phase 1-Butanol 40 % v/v 100.0 26.0 55.0 % Digital Continuous Incubation: ?, Assay: 1 M sodium acetate buffer, 10 mM NaN3, 50 Β΅M Chlorodimedone Incubation: ?, Assay: 5.5 Incubation: 30Β°C, Assay: 25Β°C 10 mM Hβ‚‚Oβ‚‚ (Hydrogen Peroxide), 100 mM Sodium bromide H2O , HOBr β€” β€” Water-incubated control (in %), assay in aqueous phase | Uncertainty about whether assay was in the presence of organic solvent or in aqueous phase
G106S/V148I Stability - Incubation Activity after incubation (1 hour at 30Β°C), measured by absorbance spectrophotometry (colorimetric assay, chlorodimedone and HOBr reaction product absorbance measurement, 290 nm) in aqueous phase 1-Butanol 30 % v/v 96.0 58.0 60.0 % Digital Continuous Incubation: ?, Assay: 1 M sodium acetate buffer, 10 mM NaN3, 50 Β΅M Chlorodimedone Incubation: ?, Assay: 5.5 Incubation: 30Β°C, Assay: 25Β°C 10 mM Hβ‚‚Oβ‚‚ (Hydrogen Peroxide), 100 mM Sodium bromide H2O , HOBr β€” β€” Water-incubated control (in %), assay in aqueous phase | Uncertainty about whether assay was in the presence of organic solvent or in aqueous phase
R114H/N146H Stability - Incubation Activity after incubation (1 hour at 30Β°C), measured by absorbance spectrophotometry (colorimetric assay, chlorodimedone and HOBr reaction product absorbance measurement, 290 nm) in aqueous phase 1-Butanol 30 % v/v 100.0 58.0 71.0 % Digital Continuous Incubation: ?, Assay: 1 M sodium acetate buffer, 10 mM NaN3, 50 Β΅M Chlorodimedone Incubation: ?, Assay: 5.5 Incubation: 30Β°C, Assay: 25Β°C 10 mM Hβ‚‚Oβ‚‚ (Hydrogen Peroxide), 100 mM Sodium bromide H2O , HOBr β€” β€” Water-incubated control (in %), assay in aqueous phase | Uncertainty about whether assay was in the presence of organic solvent or in aqueous phase
G106S/V148I Stability - Incubation Activity after incubation (1 hour at 30Β°C), measured by absorbance spectrophotometry (colorimetric assay, chlorodimedone and HOBr reaction product absorbance measurement, 290 nm) in aqueous phase 1-Butanol 20 % v/v 96.0 78.0 86.0 % Digital Continuous Incubation: ?, Assay: 1 M sodium acetate buffer, 10 mM NaN3, 50 Β΅M Chlorodimedone Incubation: ?, Assay: 5.5 Incubation: 30Β°C, Assay: 25Β°C 10 mM Hβ‚‚Oβ‚‚ (Hydrogen Peroxide), 100 mM Sodium bromide H2O , HOBr β€” β€” Water-incubated control (in %), assay in aqueous phase | Uncertainty about whether assay was in the presence of organic solvent or in aqueous phase
R114H/N146H Stability - Incubation Activity after incubation (1 hour at 30Β°C), measured by absorbance spectrophotometry (colorimetric assay, chlorodimedone and HOBr reaction product absorbance measurement, 290 nm) in aqueous phase 1-Butanol 20 % v/v 100.0 78.0 79.0 % Digital Continuous Incubation: ?, Assay: 1 M sodium acetate buffer, 10 mM NaN3, 50 Β΅M Chlorodimedone Incubation: ?, Assay: 5.5 Incubation: 30Β°C, Assay: 25Β°C 10 mM Hβ‚‚Oβ‚‚ (Hydrogen Peroxide), 100 mM Sodium bromide H2O , HOBr β€” β€” Water-incubated control (in %), assay in aqueous phase | Uncertainty about whether assay was in the presence of organic solvent or in aqueous phase
G106S/V148I Stability - Incubation Activity after incubation (1 hour at 30Β°C), measured by absorbance spectrophotometry (colorimetric assay, chlorodimedone and HOBr reaction product absorbance measurement, 290 nm) in aqueous phase 1-Butanol 10 % v/v 96.0 96.0 98.0 % Digital Continuous Incubation: ?, Assay: 1 M sodium acetate buffer, 10 mM NaN3, 50 Β΅M Chlorodimedone Incubation: ?, Assay: 5.5 Incubation: 30Β°C, Assay: 25Β°C 10 mM Hβ‚‚Oβ‚‚ (Hydrogen Peroxide), 100 mM Sodium bromide H2O , HOBr β€” β€” Water-incubated control (in %), assay in aqueous phase | Uncertainty about whether assay was in the presence of organic solvent or in aqueous phase
R114H/N146H Stability - Incubation Activity after incubation (1 hour at 30Β°C), measured by absorbance spectrophotometry (colorimetric assay, chlorodimedone and HOBr reaction product absorbance measurement, 290 nm) in aqueous phase 1-Butanol 10 % v/v 100.0 96.0 94.0 % Digital Continuous Incubation: ?, Assay: 1 M sodium acetate buffer, 10 mM NaN3, 50 Β΅M Chlorodimedone Incubation: ?, Assay: 5.5 Incubation: 30Β°C, Assay: 25Β°C 10 mM Hβ‚‚Oβ‚‚ (Hydrogen Peroxide), 100 mM Sodium bromide H2O , HOBr β€” β€” Water-incubated control (in %), assay in aqueous phase | Uncertainty about whether assay was in the presence of organic solvent or in aqueous phase
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Mutations in this dataset (7) β€” Ryosuke Yamada et al. (2014)

WT G106S G106S/V148I R114H R114H/N146H N146H V148I

Visualization : Stability β€” Incubation

Ryosuke Yamada et al. (2014)

One bar per measurement. Colour = solvent, shade = solvent volume. β€” β€” β€” Reference value. Hover for details.

Mutation Effect

Mutation impact on enzyme stability and function in the presence of organic solvent: comparison of wild-type and mutant values in identical conditions.

Ryosuke Yamada et al. (2015) β€” Random mutagenesis and selection of organic solvent-stable haloperoxidase from Streptomyces aureofaciens
Biotechnology Progress  Β· doi:10.1002/btpr.2117 β†—  Β· Stability - Incubation
34 measurements

Structure

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