Leucine dehydrogenase from Exigobacterium sibiricum

Enzyme Description

Species
Exigobacterium sibiricum β†—
Extremophile
No but psychrotolerant organism
EC Number
1.4.1.9 β†—

Sequence

Length: 373 amino acids
VETNVEARFSIFETMAMEDYEQVVFCHDKVSGLKAIIAIHDTTLGPALGGLRMWNYASDEEALIDALRLAKGMTYKKAAAGLNLGGGKAVIIGDAKTQKSEALFRAFGRYVQSLNGRYITAEDVNTTVADMDYIHMETDFVTGVSPAFGSSGNPSPVTAYGVYRGMKAAAKEVYGTDSLGGKTVAIQGVGNVAFNLCRHLHEEGAKLIVTDINQDALRRAEEAFGALVVGPDEIYSVDADIFAPCALGATLNDETIPQLKVKIIAGAANNQLKEDRHGDMLQERGILYTPDFVINAGGVINVADELDGYNRERAMKKVELVYDAVAKVIEIAKRDHLPTYRAAEKMAEERIATMGSARSQFLRRDKNILGSRG
Jana LΓΆwe et al. (2018) β€” Enantioselective synthesis of amines via reductive amination with a dehydrogenase mutant from Exigobacterium sibiricum: Substrate scope, co-solvent tolerance and biocatalyst immobilization
Bioorganic & Medicinal Chemistry  Β· doi:10.1016/j.bmc.2017.12.005 β†—  Β· Activity - Classical Activity + Stability - Incubation
9 measurements ▼
Database ID
β€”
Sequence Annotation
Explicit - Provided (2 mutations compared to WT protein)
Protein Source
Recombinant, host bacterium Escherichia coli BL21 (DE3)

Experimental Data (9 measurements)

9 measurements
Mutation Property Assay Solvent Solvent Volume Aqueous ReferenceWT Reference Measured Value Units Solution pH Temperature Substrate(s) Product(s) Cofactor(s) Shaking Comments
K77S/N270L Activity - Classical Activity measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm) in the presence of organic solvent Dimethyl Sulfoxide (DMSO) 10% 100.0 β€” 141.4 % 2 M NH4Cl buffer 9.5 30Β°C 20 mM Acetophenone, Ammonia (R)-1-Phenylethylamine 0.1 mM NADH β€” Classical aqueous control (in %)
K77S/N270L Activity - Classical Activity measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm) in the presence of organic solvent Dimethyl Sulfoxide (DMSO) 20% 100.0 β€” 185.9 % 2 M NH4Cl buffer 9.5 30Β°C 20 mM Acetophenone, Ammonia (R)-1-Phenylethylamine 0.1 mM NADH β€” Classical aqueous control (in %)
K77S/N270L Activity - Classical Activity measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm) in the presence of organic solvent Dimethyl Sulfoxide (DMSO) 30% 100.0 β€” 168.0 % 2 M NH4Cl buffer 9.5 30Β°C 20 mM Acetophenone, Ammonia (R)-1-Phenylethylamine 0.1 mM NADH β€” Classical aqueous control (in %)
K77S/N270L Activity + Stability - Incubation Activity after incubation (1 hour at 30Β°C), measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm) in the presence of organic solvent Dimethyl Sulfoxide (DMSO) 10% 56.6 β€” 159.8 % Incubation: 2 M NH4Cl buffer, Assay: 2 M NH4Cl buffer Incubation: 9.5, Assay: 9.5 Incubation: 30Β°C, Assay: 30Β°C 20 mM Acetophenone, Ammonia (R)-1-Phenylethylamine Assay: 0.1 mM NADH β€” Water-incubated control (in %) | Clear about assay in the presence of organic solvent and 100% being the assay in aqueous phase
K77S/N270L Activity + Stability - Incubation Activity after incubation (3 hours at 30Β°C), measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm) in the presence of organic solvent Dimethyl Sulfoxide (DMSO) 10% 70.2 β€” 73.6 % Incubation: 2 M NH4Cl buffer, Assay: 2 M NH4Cl buffer Incubation: 9.5, Assay: 9.5 Incubation: 30Β°C, Assay: 30Β°C 20 mM Acetophenone, Ammonia (R)-1-Phenylethylamine Assay: 0.1 mM NADH β€” Water-incubated control (in %) | Clear about assay in the presence of organic solvent and 100% being the assay in aqueous phase
K77S/N270L Activity + Stability - Incubation Activity after incubation (1 hour at 30Β°C), measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm) in the presence of organic solvent Dimethyl Sulfoxide (DMSO) 20% 56.6 β€” 173.4 % Incubation: 2 M NH4Cl buffer, Assay: 2 M NH4Cl buffer Incubation: 9.5, Assay: 9.5 Incubation: 30Β°C, Assay: 30Β°C 20 mM Acetophenone, Ammonia (R)-1-Phenylethylamine Assay: 0.1 mM NADH β€” Water-incubated control (in %) | Clear about assay in the presence of organic solvent and 100% being the assay in aqueous phase
K77S/N270L Activity + Stability - Incubation Activity after incubation (3 hours at 30Β°C), measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm) in the presence of organic solvent Dimethyl Sulfoxide (DMSO) 20% 70.2 β€” 59.6 % Incubation: 2 M NH4Cl buffer, Assay: 2 M NH4Cl buffer Incubation: 9.5, Assay: 9.5 Incubation: 30Β°C, Assay: 30Β°C 20 mM Acetophenone, Ammonia (R)-1-Phenylethylamine Assay: 0.1 mM NADH β€” Water-incubated control (in %) | Clear about assay in the presence of organic solvent and 100% being the assay in aqueous phase
K77S/N270L Activity + Stability - Incubation Activity after incubation (1 hour at 30Β°C), measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm) in the presence of organic solvent Dimethyl Sulfoxide (DMSO) 30% 56.6 β€” 150.6 % Incubation: 2 M NH4Cl buffer, Assay: 2 M NH4Cl buffer Incubation: 9.5, Assay: 9.5 Incubation: 30Β°C, Assay: 30Β°C 20 mM Acetophenone, Ammonia (R)-1-Phenylethylamine Assay: 0.1 mM NADH β€” Water-incubated control (in %) | Clear about assay in the presence of organic solvent and 100% being the assay in aqueous phase
K77S/N270L Activity + Stability - Incubation Activity after incubation (3 hours at 30Β°C), measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm) in the presence of organic solvent Dimethyl Sulfoxide (DMSO) 30% 70.2 β€” 56.2 % Incubation: 2 M NH4Cl buffer, Assay: 2 M NH4Cl buffer Incubation: 9.5, Assay: 9.5 Incubation: 30Β°C, Assay: 30Β°C 20 mM Acetophenone, Ammonia (R)-1-Phenylethylamine Assay: 0.1 mM NADH β€” Water-incubated control (in %) | Clear about assay in the presence of organic solvent and 100% being the assay in aqueous phase

Mutations in this dataset (1)

K77S/N270L

Visualization : Activity β€” Classical

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

Visualization : Activity + Stability β€” Incubation

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

Structure

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