Subtilisin BPN' from Bacillus velezensis
Enzyme Description
Sequence
MRGKKVWISLLFALALIFTMAFGSTSSAQAAGKSNGEKKYIVGFKQTMSTMSAAKKKDVISEKGGKVQKQFKYVDAASATLNEKAVKELKKDPSVAYVEEDHVAHAYAQSVPYGVSQIKAPALHSQGYTGSNVKVAVIDSGIDSSHPDLKVAGGASMVPSETNPFQDNNSHGTHVAGTVAALNNSIGVLGVAPSASLYAVKVLGADGSGQYSWIINGIEWAIANNMDVINMSLGGPSGSAALKAAVDKAVASGVVVVAAAGNEGTSGSSSTVGYPGKYPSVIAVGAVDSSNQRASFSSVGPELDVMAPGVSIQSTLPGNKYGAYNGTSMASPHVAGAAALILSKHPNWTNTQVRSSLENTTTKLGDSFYYGKGLINVQAAAQ
Keqin Chen et al. (1991)
β Enzyme Engineering for Nonaqueous Solvents. II. Additive Effects of Mutations on the Stability and Activity of Subtilisin E in Polar Organic Media
Biotechnology Progress
Β· doi:10.1021/bp00008a007 β
Β· Activity - Michaelis-Menten
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Database ID
UniProt: P00782 β
Sequence Annotation
Inferred - from protein name (106 first residues from UniProt sequence absent from the mature protein)
Protein Source
Recombinant, host bacterium Bacillus subtilis DB428
Experimental Data (6 measurements)
6 measurements
| Property | Assay | Solvent | Solvent Volume | Aqueous Reference | Measured Value | Units | Solution | pH | Temperature | Substrate(s) | Product(s) | Cofactor(s) | Shaking | Comments |
|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
| Activity - Michaelis-Menten | Km (in mM) measured by absorbance spectrophotometry (p-nitroaniline absorbance measurement, 410 nm) in the presence of organic solvent | Dimethylformamide (DMF) | 5% | 0.37 | 1.3 | mM | 0.1 MM Tris-HCl, 10 mM CaCl2 | 8 | 37Β°C | ? mM N-succinyl-Ala-Ala-Pro-Phe-p-nitroanilide | N-succinyl-Ala-Ala-Pro-Phe , p-Nitroaniline | β | β | Classical aqueous control (in mM) |
| Activity - Michaelis-Menten | Km (in mM) measured by absorbance spectrophotometry (p-nitroaniline absorbance measurement, 410 nm) in the presence of organic solvent | Dimethylformamide (DMF) | 10% | 0.37 | 2.0 | mM | 0.1 MM Tris-HCl, 10 mM CaCl2 | 8 | 37Β°C | ? mM N-succinyl-Ala-Ala-Pro-Phe-p-nitroanilide | N-succinyl-Ala-Ala-Pro-Phe , p-Nitroaniline | β | β | Classical aqueous control (in mM) |
| Activity - Michaelis-Menten | Km (in mM) measured by absorbance spectrophotometry (p-nitroaniline absorbance measurement, 410 nm) in the presence of organic solvent | Dimethylformamide (DMF) | 40% | 0.37 | 6.6 | mM | 0.1 MM Tris-HCl, 10 mM CaCl2 | 8 | 37Β°C | ? mM N-succinyl-Ala-Ala-Pro-Phe-p-nitroanilide | N-succinyl-Ala-Ala-Pro-Phe , p-Nitroaniline | β | β | Classical aqueous control (in mM) |
| Activity - Michaelis-Menten | Kcat (relative units) measured by absorbance spectrophotometry (p-nitroaniline absorbance measurement, 410 nm) in the presence of organic solvent | Dimethylformamide (DMF) | 5% | 1.0 | 1.4 | relative units, control equals 1 | 0.1 MM Tris-HCl, 10 mM CaCl2 | 8 | 37Β°C | ? mM N-succinyl-Ala-Ala-Pro-Phe-p-nitroanilide | N-succinyl-Ala-Ala-Pro-Phe , p-Nitroaniline | β | β | Classical aqueous control (in relative units, control equals 1) |
| Activity - Michaelis-Menten | Kcat (relative units) measured by absorbance spectrophotometry (p-nitroaniline absorbance measurement, 410 nm) in the presence of organic solvent | Dimethylformamide (DMF) | 10% | 1.0 | 1.1 | relative units, control equals 1 | 0.1 MM Tris-HCl, 10 mM CaCl2 | 8 | 37Β°C | ? mM N-succinyl-Ala-Ala-Pro-Phe-p-nitroanilide | N-succinyl-Ala-Ala-Pro-Phe , p-Nitroaniline | β | β | Classical aqueous control (in relative units, control equals 1) |
| Activity - Michaelis-Menten | Kcat (relative units) measured by absorbance spectrophotometry (p-nitroaniline absorbance measurement, 410 nm) in the presence of organic solvent | Dimethylformamide (DMF) | 40% | 1.0 | 0.33 | relative units, control equals 1 | 0.1 MM Tris-HCl, 10 mM CaCl2 | 8 | 37Β°C | ? mM N-succinyl-Ala-Ala-Pro-Phe-p-nitroanilide | N-succinyl-Ala-Ala-Pro-Phe , p-Nitroaniline | β | β | Classical aqueous control (in relative units, control equals 1) |
Visualization : Activity β Michaelis-Menten
One bar per measurement. Colour = solvent, shade = solvent volume.