Protease (gene sapB) from Bacillus pumilus CBS

Enzyme Description

Species
Bacillus pumilus CBS โ†—
Extremophile
No
EC Number
3.4.- โ†—

Sequence

Length: 275 amino acids
AQTVPYGIPQIKAPAVHAQGYKGANVKVAVLDTGIHAAHPDLNVAGGASFVPSEPNATQDFQSHGTHVAGTIAALDNTIGVLGVAPSASLYAVKVLDRNGDGQYSWIISGIEWAVANNMDVINMSLGGPNGSTALKNAVDTANNRGVVVVAAAGNSGSTGSTSTVGYPAKYDSTIAVANVNSSNVRNSSSSAGPELDVSAPGTSILSTVPSSGYTSYTGTSMASPHVAGAAALILSKYPNLSTTQVRQRLENTATPLGNSFYYGKGLINVQAASN
Nadia Zaraรฎ Jaouadi et al. (2012) โ€” The overexpression of the SAPB of Bacillus pumilus CBS and mutated sapB-L31I/T33S/N99Y alkaline proteases in Bacillus subtilis DB430: New attractive properties for the mutant enzyme
Bioresource Technology  ยท doi:10.1016/j.biortech.2011.11.115 โ†—  ยท Stability - Incubation
56 measurements ▼
Database ID
UniProt: A8FBN9 โ†—
Sequence Annotation
Inferred - from protein name (106 first residues from UniProt sequence absent from the mature protein)
Protein Source
Recombinant, host bacterium Bacillus subtilis DB430

Experimental Data (56 measurements)

56 measurements โ€” page 2 of 3
Mutation Property Assay Solvent Solvent Volume Aqueous ReferenceWT Reference Measured Value Units Solution pH Temperature Substrate(s) Product(s) Cofactor(s) Shaking Comments
WT Stability - Incubation Activity after incubation (24 hours at 50ยฐC), measured by absorbance spectrophotometry (colorimetric assay, Folinโ€“Ciocalteu reactant and free tyrosines reaction product absorbance measurement, 440 nm) in aqueous phase Toluene 60% 100 โ€” 114 % Incubation: ?, Assay: 100 mM Gly-NaOH Incubation: ?, Assay: 10 Incubation: 50ยฐC, Assay: 65ยฐC 1% Casein free amino acids , Peptides โ€” Incubation: 180 rpm Non-incubated control (in %), assay in aqueous phase | Uncertainty about whether assay done with or without organic solvent
WT Stability - Incubation Activity after incubation (24 hours at 50ยฐC), measured by absorbance spectrophotometry (colorimetric assay, Folinโ€“Ciocalteu reactant and free tyrosines reaction product absorbance measurement, 440 nm) in aqueous phase 1-Butanol 60% 100 โ€” 40 % Incubation: ?, Assay: 100 mM Gly-NaOH Incubation: ?, Assay: 10 Incubation: 50ยฐC, Assay: 65ยฐC 1% Casein free amino acids , Peptides โ€” Incubation: 180 rpm Non-incubated control (in %), assay in aqueous phase | Uncertainty about whether assay done with or without organic solvent
WT Stability - Incubation Activity after incubation (24 hours at 50ยฐC), measured by absorbance spectrophotometry (colorimetric assay, Folinโ€“Ciocalteu reactant and free tyrosines reaction product absorbance measurement, 440 nm) in aqueous phase Ethyl Acetate 60% 100 โ€” 70 % Incubation: ?, Assay: 100 mM Gly-NaOH Incubation: ?, Assay: 10 Incubation: 50ยฐC, Assay: 65ยฐC 1% Casein free amino acids , Peptides โ€” Incubation: 180 rpm Non-incubated control (in %), assay in aqueous phase | Uncertainty about whether assay done with or without organic solvent
WT Stability - Incubation Activity after incubation (24 hours at 50ยฐC), measured by absorbance spectrophotometry (colorimetric assay, Folinโ€“Ciocalteu reactant and free tyrosines reaction product absorbance measurement, 440 nm) in aqueous phase Isopropanol 60% 100 โ€” 105 % Incubation: ?, Assay: 100 mM Gly-NaOH Incubation: ?, Assay: 10 Incubation: 50ยฐC, Assay: 65ยฐC 1% Casein free amino acids , Peptides โ€” Incubation: 180 rpm Non-incubated control (in %), assay in aqueous phase | Uncertainty about whether assay done with or without organic solvent
WT Stability - Incubation Activity after incubation (24 hours at 50ยฐC), measured by absorbance spectrophotometry (colorimetric assay, Folinโ€“Ciocalteu reactant and free tyrosines reaction product absorbance measurement, 440 nm) in aqueous phase Acetonitrile 60% 100 โ€” 0 % Incubation: ?, Assay: 100 mM Gly-NaOH Incubation: ?, Assay: 10 Incubation: 50ยฐC, Assay: 65ยฐC 1% Casein free amino acids , Peptides โ€” Incubation: 180 rpm Non-incubated control (in %), assay in aqueous phase | Uncertainty about whether assay done with or without organic solvent
WT Stability - Incubation Activity after incubation (24 hours at 50ยฐC), measured by absorbance spectrophotometry (colorimetric assay, Folinโ€“Ciocalteu reactant and free tyrosines reaction product absorbance measurement, 440 nm) in aqueous phase Ethanol 60% 100 โ€” 70 % Incubation: ?, Assay: 100 mM Gly-NaOH Incubation: ?, Assay: 10 Incubation: 50ยฐC, Assay: 65ยฐC 1% Casein free amino acids , Peptides โ€” Incubation: 180 rpm Non-incubated control (in %), assay in aqueous phase | Uncertainty about whether assay done with or without organic solvent
WT Stability - Incubation Activity after incubation (24 hours at 50ยฐC), measured by absorbance spectrophotometry (colorimetric assay, Folinโ€“Ciocalteu reactant and free tyrosines reaction product absorbance measurement, 440 nm) in aqueous phase Methanol 60% 100 โ€” 100 % Incubation: ?, Assay: 100 mM Gly-NaOH Incubation: ?, Assay: 10 Incubation: 50ยฐC, Assay: 65ยฐC 1% Casein free amino acids , Peptides โ€” Incubation: 180 rpm Non-incubated control (in %), assay in aqueous phase | Uncertainty about whether assay done with or without organic solvent
WT Stability - Incubation Activity after incubation (24 hours at 50ยฐC), measured by absorbance spectrophotometry (colorimetric assay, Folinโ€“Ciocalteu reactant and free tyrosines reaction product absorbance measurement, 440 nm) in aqueous phase Dimethylformamide (DMF) 60% 100 โ€” 190 % Incubation: ?, Assay: 100 mM Gly-NaOH Incubation: ?, Assay: 10 Incubation: 50ยฐC, Assay: 65ยฐC 1% Casein free amino acids , Peptides โ€” Incubation: 180 rpm Non-incubated control (in %), assay in aqueous phase | Uncertainty about whether assay done with or without organic solvent
L31I/T33S/N99Y Stability - Incubation Activity after incubation (240 hours at 40ยฐC), measured by absorbance spectrophotometry (colorimetric assay, Folinโ€“Ciocalteu reactant and free tyrosines reaction product absorbance measurement, 440 nm) in aqueous phase n-Hexadecane 60% 100 101 111 % Incubation: ?, Assay: 100 mM Gly-NaOH Incubation: ?, Assay: 10 Incubation: 40ยฐC, Assay: 70ยฐC 1% Casein free amino acids , Peptides โ€” Incubation: 180 rpm Non-incubated control (in %), assay in aqueous phase | Uncertainty about whether assay done with or without organic solvent
L31I/T33S/N99Y Stability - Incubation Activity after incubation (240 hours at 40ยฐC), measured by absorbance spectrophotometry (colorimetric assay, Folinโ€“Ciocalteu reactant and free tyrosines reaction product absorbance measurement, 440 nm) in aqueous phase n-Decane 60% 100 90 107 % Incubation: ?, Assay: 100 mM Gly-NaOH Incubation: ?, Assay: 10 Incubation: 40ยฐC, Assay: 70ยฐC 1% Casein free amino acids , Peptides โ€” Incubation: 180 rpm Non-incubated control (in %), assay in aqueous phase | Uncertainty about whether assay done with or without organic solvent
L31I/T33S/N99Y Stability - Incubation Activity after incubation (240 hours at 40ยฐC), measured by absorbance spectrophotometry (colorimetric assay, Folinโ€“Ciocalteu reactant and free tyrosines reaction product absorbance measurement, 440 nm) in aqueous phase n-Octane 60% 100 106 122 % Incubation: ?, Assay: 100 mM Gly-NaOH Incubation: ?, Assay: 10 Incubation: 40ยฐC, Assay: 70ยฐC 1% Casein free amino acids , Peptides โ€” Incubation: 180 rpm Non-incubated control (in %), assay in aqueous phase | Uncertainty about whether assay done with or without organic solvent
L31I/T33S/N99Y Stability - Incubation Activity after incubation (240 hours at 40ยฐC), measured by absorbance spectrophotometry (colorimetric assay, Folinโ€“Ciocalteu reactant and free tyrosines reaction product absorbance measurement, 440 nm) in aqueous phase n-Hexane 60% 100 117 135 % Incubation: ?, Assay: 100 mM Gly-NaOH Incubation: ?, Assay: 10 Incubation: 40ยฐC, Assay: 70ยฐC 1% Casein free amino acids , Peptides โ€” Incubation: 180 rpm Non-incubated control (in %), assay in aqueous phase | Uncertainty about whether assay done with or without organic solvent
L31I/T33S/N99Y Stability - Incubation Activity after incubation (240 hours at 40ยฐC), measured by absorbance spectrophotometry (colorimetric assay, Folinโ€“Ciocalteu reactant and free tyrosines reaction product absorbance measurement, 440 nm) in aqueous phase Cyclohexane 60% 100 109 129 % Incubation: ?, Assay: 100 mM Gly-NaOH Incubation: ?, Assay: 10 Incubation: 40ยฐC, Assay: 70ยฐC 1% Casein free amino acids , Peptides โ€” Incubation: 180 rpm Non-incubated control (in %), assay in aqueous phase | Uncertainty about whether assay done with or without organic solvent
L31I/T33S/N99Y Stability - Incubation Activity after incubation (240 hours at 40ยฐC), measured by absorbance spectrophotometry (colorimetric assay, Folinโ€“Ciocalteu reactant and free tyrosines reaction product absorbance measurement, 440 nm) in aqueous phase Toluene 60% 100 93 102 % Incubation: ?, Assay: 100 mM Gly-NaOH Incubation: ?, Assay: 10 Incubation: 40ยฐC, Assay: 70ยฐC 1% Casein free amino acids , Peptides โ€” Incubation: 180 rpm Non-incubated control (in %), assay in aqueous phase | Uncertainty about whether assay done with or without organic solvent
L31I/T33S/N99Y Stability - Incubation Activity after incubation (240 hours at 40ยฐC), measured by absorbance spectrophotometry (colorimetric assay, Folinโ€“Ciocalteu reactant and free tyrosines reaction product absorbance measurement, 440 nm) in aqueous phase 1-Butanol 60% 100 25 30 % Incubation: ?, Assay: 100 mM Gly-NaOH Incubation: ?, Assay: 10 Incubation: 40ยฐC, Assay: 70ยฐC 1% Casein free amino acids , Peptides โ€” Incubation: 180 rpm Non-incubated control (in %), assay in aqueous phase | Uncertainty about whether assay done with or without organic solvent
L31I/T33S/N99Y Stability - Incubation Activity after incubation (240 hours at 40ยฐC), measured by absorbance spectrophotometry (colorimetric assay, Folinโ€“Ciocalteu reactant and free tyrosines reaction product absorbance measurement, 440 nm) in aqueous phase Ethyl Acetate 60% 100 61 78 % Incubation: ?, Assay: 100 mM Gly-NaOH Incubation: ?, Assay: 10 Incubation: 40ยฐC, Assay: 70ยฐC 1% Casein free amino acids , Peptides โ€” Incubation: 180 rpm Non-incubated control (in %), assay in aqueous phase | Uncertainty about whether assay done with or without organic solvent
L31I/T33S/N99Y Stability - Incubation Activity after incubation (240 hours at 40ยฐC), measured by absorbance spectrophotometry (colorimetric assay, Folinโ€“Ciocalteu reactant and free tyrosines reaction product absorbance measurement, 440 nm) in aqueous phase Isopropanol 60% 100 84 90 % Incubation: ?, Assay: 100 mM Gly-NaOH Incubation: ?, Assay: 10 Incubation: 40ยฐC, Assay: 70ยฐC 1% Casein free amino acids , Peptides โ€” Incubation: 180 rpm Non-incubated control (in %), assay in aqueous phase | Uncertainty about whether assay done with or without organic solvent
L31I/T33S/N99Y Stability - Incubation Activity after incubation (240 hours at 40ยฐC), measured by absorbance spectrophotometry (colorimetric assay, Folinโ€“Ciocalteu reactant and free tyrosines reaction product absorbance measurement, 440 nm) in aqueous phase Acetonitrile 60% 100 0 0 % Incubation: ?, Assay: 100 mM Gly-NaOH Incubation: ?, Assay: 10 Incubation: 40ยฐC, Assay: 70ยฐC 1% Casein free amino acids , Peptides โ€” Incubation: 180 rpm Non-incubated control (in %), assay in aqueous phase | Uncertainty about whether assay done with or without organic solvent
L31I/T33S/N99Y Stability - Incubation Activity after incubation (240 hours at 40ยฐC), measured by absorbance spectrophotometry (colorimetric assay, Folinโ€“Ciocalteu reactant and free tyrosines reaction product absorbance measurement, 440 nm) in aqueous phase Ethanol 60% 100 50 61 % Incubation: ?, Assay: 100 mM Gly-NaOH Incubation: ?, Assay: 10 Incubation: 40ยฐC, Assay: 70ยฐC 1% Casein free amino acids , Peptides โ€” Incubation: 180 rpm Non-incubated control (in %), assay in aqueous phase | Uncertainty about whether assay done with or without organic solvent
L31I/T33S/N99Y Stability - Incubation Activity after incubation (240 hours at 40ยฐC), measured by absorbance spectrophotometry (colorimetric assay, Folinโ€“Ciocalteu reactant and free tyrosines reaction product absorbance measurement, 440 nm) in aqueous phase Methanol 60% 100 75 75 % Incubation: ?, Assay: 100 mM Gly-NaOH Incubation: ?, Assay: 10 Incubation: 40ยฐC, Assay: 70ยฐC 1% Casein free amino acids , Peptides โ€” Incubation: 180 rpm Non-incubated control (in %), assay in aqueous phase | Uncertainty about whether assay done with or without organic solvent
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Mutations in this dataset (2)

WT L31I/T33S/N99Y

Visualization : Stability โ€” Incubation

One bar per measurement. Colour = solvent, shade = solvent volume. โ€” โ€” โ€” 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.

Structure

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