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 1 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
P45A/N46E/F155Y/E170K/V227A/W230F/Q252L Stability - Incubation Activity after incubation (1 hour at 4Β°C), measured by absorbance spectrophotometry (NADH absorbance measurement, 340 nm) in aqueous phase Ethanol 10% 100 β€” 103 % 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/N46E/F155Y/E170K/V227A/W230F/Q252L Stability - Incubation Activity after incubation (1 hour at 4Β°C), measured by absorbance spectrophotometry (NADH absorbance measurement, 340 nm) in aqueous phase Ethanol 20% 100 β€” 102 % 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/N46E/F155Y/E170K/V227A/W230F/Q252L Stability - Incubation Activity after incubation (1 hour at 4Β°C), measured by absorbance spectrophotometry (NADH absorbance measurement, 340 nm) in aqueous phase Ethanol 30% 100 β€” 99 % 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/N46E/F155Y/E170K/V227A/W230F/Q252L Stability - Incubation Activity after incubation (1 hour at 4Β°C), measured by absorbance spectrophotometry (NADH absorbance measurement, 340 nm) in aqueous phase Ethanol 40% 100 β€” 68 % 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/N46E/F155Y/E170K/V227A/W230F/Q252L Stability - Incubation Activity after incubation (1 hour at 4Β°C), measured by absorbance spectrophotometry (NADH absorbance measurement, 340 nm) in aqueous phase Ethanol 50% 100 β€” 22 % 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/N46E/F155Y/E170K/V227A/W230F/Q252L Stability - Incubation Activity after incubation (1 hour at 4Β°C), measured by absorbance spectrophotometry (NADH absorbance measurement, 340 nm) in aqueous phase Ethanol 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
P45A/N46E/F155Y/E170K/V227A/W230F/Q252L Stability - Incubation Activity after incubation (1 hour at 4Β°C), measured by absorbance spectrophotometry (NADH absorbance measurement, 340 nm) in aqueous phase Ethanol 70% 100 β€” 5 % 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/N46E/F155Y/E170K/V227A/W230F/Q252L Stability - Incubation Activity after incubation (1 hour at 4Β°C), measured by absorbance spectrophotometry (NADH absorbance measurement, 340 nm) in aqueous phase Ethanol 80% 100 β€” 17 % 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/N46E/F155Y/E170K/V227A/W230F/Q252L Stability - Incubation Activity after incubation (1 hour at 4Β°C), measured by absorbance spectrophotometry (NADH absorbance measurement, 340 nm) in aqueous phase Ethanol 90% 100 β€” 39 % 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/N46E/F155Y/E170K/V227A/W230F/Q252L Stability - Incubation Activity after incubation (1 hour at 4Β°C), measured by absorbance spectrophotometry (NADH absorbance measurement, 340 nm) in aqueous phase Ethanol 99% 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/Q252L Stability - Incubation Activity after incubation (1 hour at 4Β°C), measured by absorbance spectrophotometry (NADH absorbance measurement, 340 nm) in aqueous phase Ethanol 10% 100 β€” 103 % 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/Q252L Stability - Incubation Activity after incubation (1 hour at 4Β°C), measured by absorbance spectrophotometry (NADH absorbance measurement, 340 nm) in aqueous phase Ethanol 20% 100 β€” 105 % 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/Q252L Stability - Incubation Activity after incubation (1 hour at 4Β°C), measured by absorbance spectrophotometry (NADH absorbance measurement, 340 nm) in aqueous phase Ethanol 30% 100 β€” 92 % 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/Q252L Stability - Incubation Activity after incubation (1 hour at 4Β°C), measured by absorbance spectrophotometry (NADH absorbance measurement, 340 nm) in aqueous phase Ethanol 40% 100 β€” 43 % 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/Q252L Stability - Incubation Activity after incubation (1 hour at 4Β°C), measured by absorbance spectrophotometry (NADH absorbance measurement, 340 nm) in aqueous phase Ethanol 50% 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
E170K/Q252L Stability - Incubation Activity after incubation (1 hour at 4Β°C), measured by absorbance spectrophotometry (NADH absorbance measurement, 340 nm) in aqueous phase Ethanol 60% 100 β€” 5 % 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/Q252L Stability - Incubation Activity after incubation (1 hour at 4Β°C), measured by absorbance spectrophotometry (NADH absorbance measurement, 340 nm) in aqueous phase Ethanol 70% 100 β€” 19 % 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/Q252L Stability - Incubation Activity after incubation (1 hour at 4Β°C), measured by absorbance spectrophotometry (NADH absorbance measurement, 340 nm) in aqueous phase Ethanol 80% 100 β€” 41 % 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/Q252L Stability - Incubation Activity after incubation (1 hour at 4Β°C), measured by absorbance spectrophotometry (NADH absorbance measurement, 340 nm) in aqueous phase Ethanol 90% 100 β€” 71 % 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/Q252L Stability - Incubation Activity after incubation (1 hour at 4Β°C), measured by absorbance spectrophotometry (NADH absorbance measurement, 340 nm) in aqueous phase Ethanol 99% 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
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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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