3-ketosteroid-Ξ”1-dehydrogenase (gene kstD1) from Arthrobacter simplex 156

Enzyme Description

Species
Arthrobacter simplex 156 β†—
Extremophile
No
EC Number
1.3.99.4 β†—

Sequence

Length: 515 amino acids
MDWAEEYDVLVAGSGAGGMAGTYTAAREGLSVCLVEAGDKFGGTTAYSGGGGAWFPANPVLLRAGTDDTIEDALEYYRAVVGDRTPADLQETYVRGGAGLVAYLEEDDHFSFESYPWPDYFGDAPKARRDGQRHIIPTPLPVPSAPELREVVRGPLDNDRLGTPQPDDLFIGGRALVARFLTALATYPHATLVRETALAELVVEDGVVVGAIVETDGVRRAIRARRGVLLAAGGFEANDELRQKYGVPGVARDTMGPPTNVGAAHQAAIAVGADTDLMGEAWWSPGLTHPDGRSAFALWFTGGIFVDGAGRRFVNESAPYDRLGRAVIDHLTEGGVTPRYWMVYDHKEGSIPPVRATNVSMVDEEQYVAAGLWHTADTLPELAALIGVPADALVATVARFNELVADGYDADFGRGGEAYDRFFSGGEPPLVSIDEGPFHAAAFGISDLGTKGGLRTDTSARVLTADGTPIGGLYAAGNTMAAPSGTTYPGGGNPIGTSMLFSHLAVRHMGTEDAR
Shuhong Mao et al. (2018) β€” Engineering of 3-ketosteroid-βˆ†1-dehydrogenase based site-directed saturation mutagenesis for efficient biotransformation of steroidal substrates
Microbial Cell Factories  Β· doi:10.1186/s12934-018-0981-0 β†—  Β· Activity - Classical Stability - Incubation
32 measurements ▼
Database ID
UniProt: P77815 β†—
Sequence Annotation
Explicit - Provided GenBank Accession Number
Protein Source
Recombinant, host bacterium Escherichia coli BL21 (DE3)

Experimental Data (32 measurements)

32 measurements β€” 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 (2 hours at 4Β°C), measured by absorbance spectrophotometry (reduction of DCPIP to DCPIPH2, due to reduction of PMS into PMSH2 by the enzyme-FADH2, causing a decrease in absorbance, measured at 600 nm) in aqueous phase Dimethyl Sulfoxide (DMSO) 10% 100.0 157.0 % Incubation: 50 mM Tris-HCl buffer, Assay: 50 mM Tris–HCl buffer, 2% Methanol, 1.5 mM phenazine methosulphate, 40 ΞΌM 6-dichlorophenolindophenol Incubation: 8, Assay: 7.5 Incubation: 4Β°C, Assay: 30Β°C 0.5 mM Androstenedione Boldenone FAD β€” Non-incubated control (in %), assay in aqueous phase | Control with 2% methanol for subtrate solubilization
Stability - Incubation Activity after incubation (2 hours at 4Β°C), measured by absorbance spectrophotometry (reduction of DCPIP to DCPIPH2, due to reduction of PMS into PMSH2 by the enzyme-FADH2, causing a decrease in absorbance, measured at 600 nm) in aqueous phase Isopropanol 50% 100.0 110.0 % Incubation: 50 mM Tris-HCl buffer, Assay: 50 mM Tris–HCl buffer, 2% Methanol, 1.5 mM phenazine methosulphate, 40 ΞΌM 6-dichlorophenolindophenol Incubation: 8, Assay: 7.5 Incubation: 4Β°C, Assay: 30Β°C 0.5 mM Androstenedione Boldenone FAD β€” Non-incubated control (in %), assay in aqueous phase | Control with 2% methanol for subtrate solubilization
Stability - Incubation Activity after incubation (2 hours at 4Β°C), measured by absorbance spectrophotometry (reduction of DCPIP to DCPIPH2, due to reduction of PMS into PMSH2 by the enzyme-FADH2, causing a decrease in absorbance, measured at 600 nm) in aqueous phase Isopropanol 30% 100.0 124.0 % Incubation: 50 mM Tris-HCl buffer, Assay: 50 mM Tris–HCl buffer, 2% Methanol, 1.5 mM phenazine methosulphate, 40 ΞΌM 6-dichlorophenolindophenol Incubation: 8, Assay: 7.5 Incubation: 4Β°C, Assay: 30Β°C 0.5 mM Androstenedione Boldenone FAD β€” Non-incubated control (in %), assay in aqueous phase | Control with 2% methanol for subtrate solubilization
Stability - Incubation Activity after incubation (2 hours at 4Β°C), measured by absorbance spectrophotometry (reduction of DCPIP to DCPIPH2, due to reduction of PMS into PMSH2 by the enzyme-FADH2, causing a decrease in absorbance, measured at 600 nm) in aqueous phase Isopropanol 20% 100.0 133.0 % Incubation: 50 mM Tris-HCl buffer, Assay: 50 mM Tris–HCl buffer, 2% Methanol, 1.5 mM phenazine methosulphate, 40 ΞΌM 6-dichlorophenolindophenol Incubation: 8, Assay: 7.5 Incubation: 4Β°C, Assay: 30Β°C 0.5 mM Androstenedione Boldenone FAD β€” Non-incubated control (in %), assay in aqueous phase | Control with 2% methanol for subtrate solubilization
Stability - Incubation Activity after incubation (2 hours at 4Β°C), measured by absorbance spectrophotometry (reduction of DCPIP to DCPIPH2, due to reduction of PMS into PMSH2 by the enzyme-FADH2, causing a decrease in absorbance, measured at 600 nm) in aqueous phase Isopropanol 10% 100.0 126.0 % Incubation: 50 mM Tris-HCl buffer, Assay: 50 mM Tris–HCl buffer, 2% Methanol, 1.5 mM phenazine methosulphate, 40 ΞΌM 6-dichlorophenolindophenol Incubation: 8, Assay: 7.5 Incubation: 4Β°C, Assay: 30Β°C 0.5 mM Androstenedione Boldenone FAD β€” Non-incubated control (in %), assay in aqueous phase | Control with 2% methanol for subtrate solubilization
Stability - Incubation Activity after incubation (2 hours at 4Β°C), measured by absorbance spectrophotometry (reduction of DCPIP to DCPIPH2, due to reduction of PMS into PMSH2 by the enzyme-FADH2, causing a decrease in absorbance, measured at 600 nm) in aqueous phase Ethanol 50% 100.0 90.0 % Incubation: 50 mM Tris-HCl buffer, Assay: 50 mM Tris–HCl buffer, 2% Methanol, 1.5 mM phenazine methosulphate, 40 ΞΌM 6-dichlorophenolindophenol Incubation: 8, Assay: 7.5 Incubation: 4Β°C, Assay: 30Β°C 0.5 mM Androstenedione Boldenone FAD β€” Non-incubated control (in %), assay in aqueous phase | Control with 2% methanol for subtrate solubilization
Stability - Incubation Activity after incubation (2 hours at 4Β°C), measured by absorbance spectrophotometry (reduction of DCPIP to DCPIPH2, due to reduction of PMS into PMSH2 by the enzyme-FADH2, causing a decrease in absorbance, measured at 600 nm) in aqueous phase Ethanol 30% 100.0 128.0 % Incubation: 50 mM Tris-HCl buffer, Assay: 50 mM Tris–HCl buffer, 2% Methanol, 1.5 mM phenazine methosulphate, 40 ΞΌM 6-dichlorophenolindophenol Incubation: 8, Assay: 7.5 Incubation: 4Β°C, Assay: 30Β°C 0.5 mM Androstenedione Boldenone FAD β€” Non-incubated control (in %), assay in aqueous phase | Control with 2% methanol for subtrate solubilization
Stability - Incubation Activity after incubation (2 hours at 4Β°C), measured by absorbance spectrophotometry (reduction of DCPIP to DCPIPH2, due to reduction of PMS into PMSH2 by the enzyme-FADH2, causing a decrease in absorbance, measured at 600 nm) in aqueous phase Ethanol 20% 100.0 137.0 % Incubation: 50 mM Tris-HCl buffer, Assay: 50 mM Tris–HCl buffer, 2% Methanol, 1.5 mM phenazine methosulphate, 40 ΞΌM 6-dichlorophenolindophenol Incubation: 8, Assay: 7.5 Incubation: 4Β°C, Assay: 30Β°C 0.5 mM Androstenedione Boldenone FAD β€” Non-incubated control (in %), assay in aqueous phase | Control with 2% methanol for subtrate solubilization
Stability - Incubation Activity after incubation (2 hours at 4Β°C), measured by absorbance spectrophotometry (reduction of DCPIP to DCPIPH2, due to reduction of PMS into PMSH2 by the enzyme-FADH2, causing a decrease in absorbance, measured at 600 nm) in aqueous phase Ethanol 10% 100.0 132.0 % Incubation: 50 mM Tris-HCl buffer, Assay: 50 mM Tris–HCl buffer, 2% Methanol, 1.5 mM phenazine methosulphate, 40 ΞΌM 6-dichlorophenolindophenol Incubation: 8, Assay: 7.5 Incubation: 4Β°C, Assay: 30Β°C 0.5 mM Androstenedione Boldenone FAD β€” Non-incubated control (in %), assay in aqueous phase | Control with 2% methanol for subtrate solubilization
Stability - Incubation Activity after incubation (2 hours at 4Β°C), measured by absorbance spectrophotometry (reduction of DCPIP to DCPIPH2, due to reduction of PMS into PMSH2 by the enzyme-FADH2, causing a decrease in absorbance, measured at 600 nm) in aqueous phase Methanol 50% 100.0 59.0 % Incubation: 50 mM Tris-HCl buffer, Assay: 50 mM Tris–HCl buffer, 2% Methanol, 1.5 mM phenazine methosulphate, 40 ΞΌM 6-dichlorophenolindophenol Incubation: 8, Assay: 7.5 Incubation: 4Β°C, Assay: 30Β°C 0.5 mM Androstenedione Boldenone FAD β€” Non-incubated control (in %), assay in aqueous phase | Control with 2% methanol for subtrate solubilization
Stability - Incubation Activity after incubation (2 hours at 4Β°C), measured by absorbance spectrophotometry (reduction of DCPIP to DCPIPH2, due to reduction of PMS into PMSH2 by the enzyme-FADH2, causing a decrease in absorbance, measured at 600 nm) in aqueous phase Methanol 30% 100.0 121.0 % Incubation: 50 mM Tris-HCl buffer, Assay: 50 mM Tris–HCl buffer, 2% Methanol, 1.5 mM phenazine methosulphate, 40 ΞΌM 6-dichlorophenolindophenol Incubation: 8, Assay: 7.5 Incubation: 4Β°C, Assay: 30Β°C 0.5 mM Androstenedione Boldenone FAD β€” Non-incubated control (in %), assay in aqueous phase | Control with 2% methanol for subtrate solubilization
Stability - Incubation Activity after incubation (2 hours at 4Β°C), measured by absorbance spectrophotometry (reduction of DCPIP to DCPIPH2, due to reduction of PMS into PMSH2 by the enzyme-FADH2, causing a decrease in absorbance, measured at 600 nm) in aqueous phase Methanol 20% 100.0 133.0 % Incubation: 50 mM Tris-HCl buffer, Assay: 50 mM Tris–HCl buffer, 2% Methanol, 1.5 mM phenazine methosulphate, 40 ΞΌM 6-dichlorophenolindophenol Incubation: 8, Assay: 7.5 Incubation: 4Β°C, Assay: 30Β°C 0.5 mM Androstenedione Boldenone FAD β€” Non-incubated control (in %), assay in aqueous phase | Control with 2% methanol for subtrate solubilization
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Visualization : Activity β€” Classical

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

Visualization : Stability β€” Incubation

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

Structure

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