Glucose 1-dehydrogenase from Bacillus subtilis 168

Enzyme Description

Species
Bacillus subtilis 168 β†—
Extremophile
No
EC Number
1.1.1.47 β†—

Sequence

Length: 261 amino acids
MYPDLKGKVVAITGAASGLGKAMAIRFGKEQAKVVINYYSNKQDPNEVKEEVIKAGGEAVVVQGDVTKEEDVKNIVQTAIKEFGTLDIMINNAGLENPVPSHEMPLKDWDKVIGTNLTGAFLGSREAIKYFVENDIKGNVINMSSVHEVIPWPLFVHYAASKGGIKLMTETLALEYAPKGIRVNNIGPGAINTPINAEKFADPKQKADVESMIPMGYIGEPEEIAAVAAWLASKEASYVTGITLFADGGMTQYPSFQAGRG
E. Vazquez-Figueroa et al. (2008) β€” Thermostable variants constructed via the structure-guided consensus method also show increased stability in salts solutions and homogeneous aqueous-organic media
Protein Engineering, Design and Selection  Β· doi:10.1093/protein/gzn048 β†—  Β· Stability - Incubation
120 measurements ▼
Database ID
UniProt: P12310 β†—
Sequence Annotation
Inferred - from protein name
Protein Source
Recombinant, host bacterium Escherichia coli BL21 (DE3)

Experimental Data (120 measurements)

120 measurements β€” page 4 of 6
Mutation Property Assay Solvent Solvent Volume Aqueous ReferenceWT Reference Measured Value Units Solution pH Temperature Substrate(s) Product(s) Cofactor(s) Shaking Comments
E170K Stability - Incubation Activity after incubation (1 hour at 4Β°C), measured by absorbance spectrophotometry (NADH absorbance measurement, 340 nm) in aqueous phase Acetone 10% 100 β€” 89 % Incubation: 5 mM sodium phosphate buffer, Assay: 5 mM sodium phosphate buffer Incubation: 8, Assay: 8 Incubation: 4Β°C, Assay: 4Β°C 100 mM D-Glucose D-glucono-1,5-lactone 1 mM NAD+ β€” Water-incubated control (in %), assay in aqueous phase | Clear about assay in aqueous phase
E170K Stability - Incubation Activity after incubation (1 hour at 4Β°C), measured by absorbance spectrophotometry (NADH absorbance measurement, 340 nm) in aqueous phase Acetone 20% 100 β€” 63 % Incubation: 5 mM sodium phosphate buffer, Assay: 5 mM sodium phosphate buffer Incubation: 8, Assay: 8 Incubation: 4Β°C, Assay: 4Β°C 100 mM D-Glucose D-glucono-1,5-lactone 1 mM NAD+ β€” Water-incubated control (in %), assay in aqueous phase | Clear about assay in aqueous phase
E170K Stability - Incubation Activity after incubation (1 hour at 4Β°C), measured by absorbance spectrophotometry (NADH absorbance measurement, 340 nm) in aqueous phase Acetone 30% 100 β€” 31 % Incubation: 5 mM sodium phosphate buffer, Assay: 5 mM sodium phosphate buffer Incubation: 8, Assay: 8 Incubation: 4Β°C, Assay: 4Β°C 100 mM D-Glucose D-glucono-1,5-lactone 1 mM NAD+ β€” Water-incubated control (in %), assay in aqueous phase | Clear about assay in aqueous phase
E170K Stability - Incubation Activity after incubation (1 hour at 4Β°C), measured by absorbance spectrophotometry (NADH absorbance measurement, 340 nm) in aqueous phase Acetone 40% 100 β€” 0 % Incubation: 5 mM sodium phosphate buffer, Assay: 5 mM sodium phosphate buffer Incubation: 8, Assay: 8 Incubation: 4Β°C, Assay: 4Β°C 100 mM D-Glucose D-glucono-1,5-lactone 1 mM NAD+ β€” Water-incubated control (in %), assay in aqueous phase | Clear about assay in aqueous phase
E170K Stability - Incubation Activity after incubation (1 hour at 4Β°C), measured by absorbance spectrophotometry (NADH absorbance measurement, 340 nm) in aqueous phase Acetone 50% 100 β€” 0 % Incubation: 5 mM sodium phosphate buffer, Assay: 5 mM sodium phosphate buffer Incubation: 8, Assay: 8 Incubation: 4Β°C, Assay: 4Β°C 100 mM D-Glucose D-glucono-1,5-lactone 1 mM NAD+ β€” Water-incubated control (in %), assay in aqueous phase | Clear about assay in aqueous phase
E170K Stability - Incubation Activity after incubation (1 hour at 4Β°C), measured by absorbance spectrophotometry (NADH absorbance measurement, 340 nm) in aqueous phase Acetone 60% 100 β€” 1 % Incubation: 5 mM sodium phosphate buffer, Assay: 5 mM sodium phosphate buffer Incubation: 8, Assay: 8 Incubation: 4Β°C, Assay: 4Β°C 100 mM D-Glucose D-glucono-1,5-lactone 1 mM NAD+ β€” Water-incubated control (in %), assay in aqueous phase | Clear about assay in aqueous phase
E170K Stability - Incubation Activity after incubation (1 hour at 4Β°C), measured by absorbance spectrophotometry (NADH absorbance measurement, 340 nm) in aqueous phase Acetone 70% 100 β€” 12 % Incubation: 5 mM sodium phosphate buffer, Assay: 5 mM sodium phosphate buffer Incubation: 8, Assay: 8 Incubation: 4Β°C, Assay: 4Β°C 100 mM D-Glucose D-glucono-1,5-lactone 1 mM NAD+ β€” Water-incubated control (in %), assay in aqueous phase | Clear about assay in aqueous phase
E170K Stability - Incubation Activity after incubation (1 hour at 4Β°C), measured by absorbance spectrophotometry (NADH absorbance measurement, 340 nm) in aqueous phase Acetone 80% 100 β€” 25 % Incubation: 5 mM sodium phosphate buffer, Assay: 5 mM sodium phosphate buffer Incubation: 8, Assay: 8 Incubation: 4Β°C, Assay: 4Β°C 100 mM D-Glucose D-glucono-1,5-lactone 1 mM NAD+ β€” Water-incubated control (in %), assay in aqueous phase | Clear about assay in aqueous phase
E170K Stability - Incubation Activity after incubation (1 hour at 4Β°C), measured by absorbance spectrophotometry (NADH absorbance measurement, 340 nm) in aqueous phase Acetone 90% 100 β€” 42 % Incubation: 5 mM sodium phosphate buffer, Assay: 5 mM sodium phosphate buffer Incubation: 8, Assay: 8 Incubation: 4Β°C, Assay: 4Β°C 100 mM D-Glucose D-glucono-1,5-lactone 1 mM NAD+ β€” Water-incubated control (in %), assay in aqueous phase | Clear about assay in aqueous phase
E170K Stability - Incubation Activity after incubation (1 hour at 4Β°C), measured by absorbance spectrophotometry (NADH absorbance measurement, 340 nm) in aqueous phase Acetone 99% 100 β€” 23 % Incubation: 5 mM sodium phosphate buffer, Assay: 5 mM sodium phosphate buffer Incubation: 8, Assay: 8 Incubation: 4Β°C, Assay: 4Β°C 100 mM D-Glucose D-glucono-1,5-lactone 1 mM NAD+ β€” Water-incubated control (in %), assay in aqueous phase | Clear about assay in aqueous phase
P45A/F155Y/V227A Stability - Incubation Activity after incubation (1 hour at 4Β°C), measured by absorbance spectrophotometry (NADH absorbance measurement, 340 nm) in aqueous phase Acetone 10% 100 β€” 40 % Incubation: 5 mM sodium phosphate buffer, Assay: 5 mM sodium phosphate buffer Incubation: 8, Assay: 8 Incubation: 4Β°C, Assay: 4Β°C 100 mM D-Glucose D-glucono-1,5-lactone 1 mM NAD+ β€” Water-incubated control (in %), assay in aqueous phase | Clear about assay in aqueous phase
P45A/F155Y/V227A Stability - Incubation Activity after incubation (1 hour at 4Β°C), measured by absorbance spectrophotometry (NADH absorbance measurement, 340 nm) in aqueous phase Acetone 20% 100 β€” 6 % Incubation: 5 mM sodium phosphate buffer, Assay: 5 mM sodium phosphate buffer Incubation: 8, Assay: 8 Incubation: 4Β°C, Assay: 4Β°C 100 mM D-Glucose D-glucono-1,5-lactone 1 mM NAD+ β€” Water-incubated control (in %), assay in aqueous phase | Clear about assay in aqueous phase
P45A/F155Y/V227A Stability - Incubation Activity after incubation (1 hour at 4Β°C), measured by absorbance spectrophotometry (NADH absorbance measurement, 340 nm) in aqueous phase Acetone 30% 100 β€” 0 % Incubation: 5 mM sodium phosphate buffer, Assay: 5 mM sodium phosphate buffer Incubation: 8, Assay: 8 Incubation: 4Β°C, Assay: 4Β°C 100 mM D-Glucose D-glucono-1,5-lactone 1 mM NAD+ β€” Water-incubated control (in %), assay in aqueous phase | Clear about assay in aqueous phase
P45A/F155Y/V227A Stability - Incubation Activity after incubation (1 hour at 4Β°C), measured by absorbance spectrophotometry (NADH absorbance measurement, 340 nm) in aqueous phase Acetone 40% 100 β€” 0 % Incubation: 5 mM sodium phosphate buffer, Assay: 5 mM sodium phosphate buffer Incubation: 8, Assay: 8 Incubation: 4Β°C, Assay: 4Β°C 100 mM D-Glucose D-glucono-1,5-lactone 1 mM NAD+ β€” Water-incubated control (in %), assay in aqueous phase | Clear about assay in aqueous phase
P45A/F155Y/V227A Stability - Incubation Activity after incubation (1 hour at 4Β°C), measured by absorbance spectrophotometry (NADH absorbance measurement, 340 nm) in aqueous phase Acetone 50% 100 β€” 0 % Incubation: 5 mM sodium phosphate buffer, Assay: 5 mM sodium phosphate buffer Incubation: 8, Assay: 8 Incubation: 4Β°C, Assay: 4Β°C 100 mM D-Glucose D-glucono-1,5-lactone 1 mM NAD+ β€” Water-incubated control (in %), assay in aqueous phase | Clear about assay in aqueous phase
P45A/F155Y/V227A Stability - Incubation Activity after incubation (1 hour at 4Β°C), measured by absorbance spectrophotometry (NADH absorbance measurement, 340 nm) in aqueous phase Acetone 60% 100 β€” 0 % Incubation: 5 mM sodium phosphate buffer, Assay: 5 mM sodium phosphate buffer Incubation: 8, Assay: 8 Incubation: 4Β°C, Assay: 4Β°C 100 mM D-Glucose D-glucono-1,5-lactone 1 mM NAD+ β€” Water-incubated control (in %), assay in aqueous phase | Clear about assay in aqueous phase
P45A/F155Y/V227A Stability - Incubation Activity after incubation (1 hour at 4Β°C), measured by absorbance spectrophotometry (NADH absorbance measurement, 340 nm) in aqueous phase Acetone 70% 100 β€” 0 % Incubation: 5 mM sodium phosphate buffer, Assay: 5 mM sodium phosphate buffer Incubation: 8, Assay: 8 Incubation: 4Β°C, Assay: 4Β°C 100 mM D-Glucose D-glucono-1,5-lactone 1 mM NAD+ β€” Water-incubated control (in %), assay in aqueous phase | Clear about assay in aqueous phase
P45A/F155Y/V227A Stability - Incubation Activity after incubation (1 hour at 4Β°C), measured by absorbance spectrophotometry (NADH absorbance measurement, 340 nm) in aqueous phase Acetone 80% 100 β€” 1 % Incubation: 5 mM sodium phosphate buffer, Assay: 5 mM sodium phosphate buffer Incubation: 8, Assay: 8 Incubation: 4Β°C, Assay: 4Β°C 100 mM D-Glucose D-glucono-1,5-lactone 1 mM NAD+ β€” Water-incubated control (in %), assay in aqueous phase | Clear about assay in aqueous phase
P45A/F155Y/V227A Stability - Incubation Activity after incubation (1 hour at 4Β°C), measured by absorbance spectrophotometry (NADH absorbance measurement, 340 nm) in aqueous phase Acetone 90% 100 β€” 8 % Incubation: 5 mM sodium phosphate buffer, Assay: 5 mM sodium phosphate buffer Incubation: 8, Assay: 8 Incubation: 4Β°C, Assay: 4Β°C 100 mM D-Glucose D-glucono-1,5-lactone 1 mM NAD+ β€” Water-incubated control (in %), assay in aqueous phase | Clear about assay in aqueous phase
P45A/F155Y/V227A Stability - Incubation Activity after incubation (1 hour at 4Β°C), measured by absorbance spectrophotometry (NADH absorbance measurement, 340 nm) in aqueous phase Acetone 99% 100 β€” 31 % Incubation: 5 mM sodium phosphate buffer, Assay: 5 mM sodium phosphate buffer Incubation: 8, Assay: 8 Incubation: 4Β°C, Assay: 4Β°C 100 mM D-Glucose D-glucono-1,5-lactone 1 mM NAD+ β€” Water-incubated control (in %), assay in aqueous phase | Clear about assay in aqueous phase
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Mutations in this dataset (4)

P45A/F155Y/V227A P45A/N46E/F155Y/E170K/V227A/W230F/Q252L E170K E170K/Q252L

Visualization : Stability β€” Incubation

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

Structure

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