Vimelysin from Vibrio sp. T-1800
Enzyme Description
Sequence
AKSSGTGPGGNLKTTRYEYGSDFPSFSIDKTGTTCKLENDSVKTVNLNHGTSGSAAYSYNCADGTNYTDHKYINGAYSPLNDAHYFGNVVFDMYKEWMNTSPLTFQLTMRVHYSSNYENAFWNGSSMTFGDGGSTFYPLVDINVSAHEVSHGFTEQNSGLVYQNMSGGINEAFSDIAGEAAEYYLRGNVDWVVGSDIFKSEGGLRYFDQPSKDGRSIDHASQYYDGLNVHLSSGVYNRAFYLLANKSGWDVRKGFEIFTVANQLYWTANSTFDAGACGVAKAAADMGYVVADVEDAFNTVGVNGSCGSTPPTGNVLTKGTPIANLSGNQSSESFYTFTVDSASSATVSMSGGSGDADLYVKSGSKPTTSSYDCRPYRAGNNEQCSVSAQPGITYHVLLRGYSNYSGLTLRLD
Kohei Oda et al. (1996)
β A Novel Alcohol Resistant Metalloproteinase, Vimelysin, from Vibrio sp. T1800: Purification and Characterization
Bioscience, Biotechnology, and Biochemistry
Β· doi:10.1271/bbb.60.463 β
Β· Activity - Classical
▼
Database ID
UniProt: Q76LC2 β
Sequence Annotation
Inferred - from protein name
Protein Source
Purified, bacterium Vibrio sp. T-1800
Experimental Data (22 measurements)
22 measurements β page 1 of 2
| Property | Assay | Solvent | Solvent Volume | Aqueous Reference | Measured Value | Units | Solution | pH | Temperature | Substrate(s) | Product(s) | Cofactor(s) | Shaking | Comments |
|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
| Activity - Classical | Activity measured by absorbance spectrophotometry (FAGLA absorbance measurement, 345 nm) in the presence of organic solvent | Isopropanol | 10% | 100 | 14 | % | 0.1 M MES | 6.5 | ? | 1 mM FAGLA (FA-Gly-DL-Leu-NH2) | Furylacryloyl-Gly , Leu-NH2 | β | β | Classical aqueous control (in %) |
| Activity - Classical | Activity measured by absorbance spectrophotometry (FAGLA absorbance measurement, 345 nm) in the presence of organic solvent | Dimethylformamide (DMF) | 30% | 100 | 1 | % | 0.1 M MES | 6.5 | ? | 1 mM FAGLA (FA-Gly-DL-Leu-NH2) | Furylacryloyl-Gly , Leu-NH2 | β | β | Classical aqueous control (in %) |
| Activity - Classical | Activity measured by absorbance spectrophotometry (FAGLA absorbance measurement, 345 nm) in the presence of organic solvent | Dimethylformamide (DMF) | 20% | 100 | 10 | % | 0.1 M MES | 6.5 | ? | 1 mM FAGLA (FA-Gly-DL-Leu-NH2) | Furylacryloyl-Gly , Leu-NH2 | β | β | Classical aqueous control (in %) |
| Activity - Classical | Activity measured by absorbance spectrophotometry (FAGLA absorbance measurement, 345 nm) in the presence of organic solvent | Dimethylformamide (DMF) | 10% | 100 | 26 | % | 0.1 M MES | 6.5 | ? | 1 mM FAGLA (FA-Gly-DL-Leu-NH2) | Furylacryloyl-Gly , Leu-NH2 | β | β | Classical aqueous control (in %) |
| Activity - Classical | Activity measured by absorbance spectrophotometry (FAGLA absorbance measurement, 345 nm) in the presence of organic solvent | Dimethylformamide (DMF) | 5% | 100 | 39 | % | 0.1 M MES | 6.5 | ? | 1 mM FAGLA (FA-Gly-DL-Leu-NH2) | Furylacryloyl-Gly , Leu-NH2 | β | β | Classical aqueous control (in %) |
| Activity - Classical | Activity measured by absorbance spectrophotometry (FAGLA absorbance measurement, 345 nm) in the presence of organic solvent | Dimethyl Sulfoxide (DMSO) | 30% | 100 | 28 | % | 0.1 M MES | 6.5 | ? | 1 mM FAGLA (FA-Gly-DL-Leu-NH2) | Furylacryloyl-Gly , Leu-NH2 | β | β | Classical aqueous control (in %) |
| Activity - Classical | Activity measured by absorbance spectrophotometry (FAGLA absorbance measurement, 345 nm) in the presence of organic solvent | Dimethyl Sulfoxide (DMSO) | 20% | 100 | 46 | % | 0.1 M MES | 6.5 | ? | 1 mM FAGLA (FA-Gly-DL-Leu-NH2) | Furylacryloyl-Gly , Leu-NH2 | β | β | Classical aqueous control (in %) |
| Activity - Classical | Activity measured by absorbance spectrophotometry (FAGLA absorbance measurement, 345 nm) in the presence of organic solvent | Dimethyl Sulfoxide (DMSO) | 10% | 100 | 70 | % | 0.1 M MES | 6.5 | ? | 1 mM FAGLA (FA-Gly-DL-Leu-NH2) | Furylacryloyl-Gly , Leu-NH2 | β | β | Classical aqueous control (in %) |
| Activity - Classical | Activity measured by absorbance spectrophotometry (FAGLA absorbance measurement, 345 nm) in the presence of organic solvent | Dimethyl Sulfoxide (DMSO) | 5% | 100 | 88 | % | 0.1 M MES | 6.5 | ? | 1 mM FAGLA (FA-Gly-DL-Leu-NH2) | Furylacryloyl-Gly , Leu-NH2 | β | β | Classical aqueous control (in %) |
| Activity - Classical | Activity measured by absorbance spectrophotometry (FAGLA absorbance measurement, 345 nm) in the presence of organic solvent | Isopropanol | 30% | 100 | 1 | % | 0.1 M MES | 6.5 | ? | 1 mM FAGLA (FA-Gly-DL-Leu-NH2) | Furylacryloyl-Gly , Leu-NH2 | β | β | Classical aqueous control (in %) |
| Activity - Classical | Activity measured by absorbance spectrophotometry (FAGLA absorbance measurement, 345 nm) in the presence of organic solvent | Isopropanol | 20% | 100 | 4 | % | 0.1 M MES | 6.5 | ? | 1 mM FAGLA (FA-Gly-DL-Leu-NH2) | Furylacryloyl-Gly , Leu-NH2 | β | β | Classical aqueous control (in %) |
| Activity - Classical | Activity measured by absorbance spectrophotometry (FAGLA absorbance measurement, 345 nm) in the presence of organic solvent | Methanol | 10% | 100 | 61 | % | 0.1 M MES | 6.5 | ? | 1 mM FAGLA (FA-Gly-DL-Leu-NH2) | Furylacryloyl-Gly , Leu-NH2 | β | β | Classical aqueous control (in %) |
| Activity - Classical | Activity measured by absorbance spectrophotometry (FAGLA absorbance measurement, 345 nm) in the presence of organic solvent | Isopropanol | 5% | 100 | 25 | % | 0.1 M MES | 6.5 | ? | 1 mM FAGLA (FA-Gly-DL-Leu-NH2) | Furylacryloyl-Gly , Leu-NH2 | β | β | Classical aqueous control (in %) |
| Activity - Classical | Activity measured by absorbance spectrophotometry (FAGLA absorbance measurement, 345 nm) in the presence of organic solvent | Isopropanol | 30% | 100 | 2 | % | 0.1 M MES | 6.5 | ? | 1 mM FAGLA (FA-Gly-DL-Leu-NH2) | Furylacryloyl-Gly , Leu-NH2 | β | β | Classical aqueous control (in %) |
| Activity - Classical | Activity measured by absorbance spectrophotometry (FAGLA absorbance measurement, 345 nm) in the presence of organic solvent | Isopropanol | 20% | 100 | 16 | % | 0.1 M MES | 6.5 | ? | 1 mM FAGLA (FA-Gly-DL-Leu-NH2) | Furylacryloyl-Gly , Leu-NH2 | β | β | Classical aqueous control (in %) |
| Activity - Classical | Activity measured by absorbance spectrophotometry (FAGLA absorbance measurement, 345 nm) in the presence of organic solvent | Isopropanol | 10% | 100 | 21 | % | 0.1 M MES | 6.5 | ? | 1 mM FAGLA (FA-Gly-DL-Leu-NH2) | Furylacryloyl-Gly , Leu-NH2 | β | β | Classical aqueous control (in %) |
| Activity - Classical | Activity measured by absorbance spectrophotometry (FAGLA absorbance measurement, 345 nm) in the presence of organic solvent | Isopropanol | 5% | 100 | 35 | % | 0.1 M MES | 6.5 | ? | 1 mM FAGLA (FA-Gly-DL-Leu-NH2) | Furylacryloyl-Gly , Leu-NH2 | β | β | Classical aqueous control (in %) |
| Activity - Classical | Activity measured by absorbance spectrophotometry (FAGLA absorbance measurement, 345 nm) in the presence of organic solvent | Ethanol | 30% | 100 | 11 | % | 0.1 M MES | 6.5 | ? | 1 mM FAGLA (FA-Gly-DL-Leu-NH2) | Furylacryloyl-Gly , Leu-NH2 | β | β | Classical aqueous control (in %) |
| Activity - Classical | Activity measured by absorbance spectrophotometry (FAGLA absorbance measurement, 345 nm) in the presence of organic solvent | Ethanol | 20% | 100 | 18 | % | 0.1 M MES | 6.5 | ? | 1 mM FAGLA (FA-Gly-DL-Leu-NH2) | Furylacryloyl-Gly , Leu-NH2 | β | β | Classical aqueous control (in %) |
| Activity - Classical | Activity measured by absorbance spectrophotometry (FAGLA absorbance measurement, 345 nm) in the presence of organic solvent | Ethanol | 10% | 100 | 42 | % | 0.1 M MES | 6.5 | ? | 1 mM FAGLA (FA-Gly-DL-Leu-NH2) | Furylacryloyl-Gly , Leu-NH2 | β | β | Classical aqueous control (in %) |
Visualization : Activity β Classical
Kohei Oda et al. (1996)
One bar per measurement. Colour = solvent, shade = solvent volume.
Toshihiro Takahashi et al. (2005)
β Molecular Cloning of the Gene Encoding Vibrio Metalloproteinase Vimelysin and Isolation of a Mutant with High Stability in Organic Solvents
The Journal of Biochemistry
Β· doi:10.1093/jb/mvi173 β
Β· Stability - Incubation
▼
Database ID
UniProt: Q76LC2 β
Sequence Annotation
Explicit - Provided (first 195 residues from UniProt sequence absent from the mature protein)
Protein Source
Recombinant, host bacterium Escherichia coli JM109
Experimental Data (8 measurements)
8 measurements
| Mutation | Property | Assay | Solvent | Solvent Volume | Aqueous Reference | WT Reference | Measured Value | Units | Solution | pH | Temperature | Substrate(s) | Product(s) | Cofactor(s) | Shaking | Comments |
|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
| WT | Stability - Incubation | Activity after incubation (1 hour at 37Β°C), measured by fluorescence spectrophotometry (fluorophore MOCAc fluorescence measurement as it is separated from quencher DNP, excitation at 360 nm and absorbance measurement at 450 nm) in aqueous phase | Ethanol | 50% | 98 | β | 58 | % | Incubation: 50 mM Tris-Hcl, 5mM CaCl2, Assay: 50 mM MES buffer, 10 mM CaCl2 and 0.005% Triton X-100 | Incubation: 7.5, Assay: 6.5 | Incubation: 37Β°C, Assay: 25Β°C | MOCAc-PLGL-A2pr(Dnp)-AR-NH2 | L-A2pr(Dnp)-AR-NH2 , βMOCAc-PLG | β | β | Non-incubated control (in %), assay in aqueous phase |
| WT | Stability - Incubation | Activity after incubation (3 hours at 37Β°C), measured by fluorescence spectrophotometry (fluorophore MOCAc fluorescence measurement as it is separated from quencher DNP, excitation at 360 nm and absorbance measurement at 450 nm) in aqueous phase | Ethanol | 50% | 91 | β | 22 | % | Incubation: 50 mM Tris-Hcl, 5mM CaCl2, Assay: 50 mM MES buffer, 10 mM CaCl2 and 0.005% Triton X-100 | Incubation: 7.5, Assay: 6.5 | Incubation: 37Β°C, Assay: 25Β°C | MOCAc-PLGL-A2pr(Dnp)-AR-NH2 | L-A2pr(Dnp)-AR-NH2 , βMOCAc-PLG | β | β | Non-incubated control (in %), assay in aqueous phase |
| WT | Stability - Incubation | Activity after incubation (1 hour at 37Β°C), measured by fluorescence spectrophotometry (fluorophore MOCAc fluorescence measurement as it is separated from quencher DNP, excitation at 360 nm and absorbance measurement at 450 nm) in aqueous phase | Isopropanol | 50% | 98 | β | 72 | % | Incubation: 50 mM Tris-Hcl, 5mM CaCl2, Assay: 50 mM MES buffer, 10 mM CaCl2 and 0.005% Triton X-100 | Incubation: 7.5, Assay: 6.5 | Incubation: 37Β°C, Assay: 25Β°C | MOCAc-PLGL-A2pr(Dnp)-AR-NH2 | L-A2pr(Dnp)-AR-NH2 , βMOCAc-PLG | β | β | Non-incubated control (in %), assay in aqueous phase |
| WT | Stability - Incubation | Activity after incubation (3 hours at 37Β°C), measured by fluorescence spectrophotometry (fluorophore MOCAc fluorescence measurement as it is separated from quencher DNP, excitation at 360 nm and absorbance measurement at 450 nm) in aqueous phase | Isopropanol | 50% | 91 | β | 38 | % | Incubation: 50 mM Tris-Hcl, 5mM CaCl2, Assay: 50 mM MES buffer, 10 mM CaCl2 and 0.005% Triton X-100 | Incubation: 7.5, Assay: 6.5 | Incubation: 37Β°C, Assay: 25Β°C | MOCAc-PLGL-A2pr(Dnp)-AR-NH2 | L-A2pr(Dnp)-AR-NH2 , βMOCAc-PLG | β | β | Non-incubated control (in %), assay in aqueous phase |
| N123D | Stability - Incubation | Activity after incubation (1 hour at 37Β°C), measured by fluorescence spectrophotometry (fluorophore MOCAc fluorescence measurement as it is separated from quencher DNP, excitation at 360 nm and absorbance measurement at 450 nm) in aqueous phase | Ethanol | 50% | 102 | 58 | 79 | % | Incubation: 50 mM Tris-Hcl, 5mM CaCl2, Assay: 50 mM MES buffer, 10 mM CaCl2 and 0.005% Triton X-100 | Incubation: 7.5, Assay: 6.5 | Incubation: 37Β°C, Assay: 25Β°C | MOCAc-PLGL-A2pr(Dnp)-AR-NH2 | L-A2pr(Dnp)-AR-NH2 , βMOCAc-PLG | β | β | Non-incubated control (in %), assay in aqueous phase |
| N123D | Stability - Incubation | Activity after incubation (3 hours at 37Β°C), measured by fluorescence spectrophotometry (fluorophore MOCAc fluorescence measurement as it is separated from quencher DNP, excitation at 360 nm and absorbance measurement at 450 nm) in aqueous phase | Ethanol | 50% | 92 | 22 | 56 | % | Incubation: 50 mM Tris-Hcl, 5mM CaCl2, Assay: 50 mM MES buffer, 10 mM CaCl2 and 0.005% Triton X-100 | Incubation: 7.5, Assay: 6.5 | Incubation: 37Β°C, Assay: 25Β°C | MOCAc-PLGL-A2pr(Dnp)-AR-NH2 | L-A2pr(Dnp)-AR-NH2 , βMOCAc-PLG | β | β | Non-incubated control (in %), assay in aqueous phase |
| N123D | Stability - Incubation | Activity after incubation (1 hour at 37Β°C), measured by fluorescence spectrophotometry (fluorophore MOCAc fluorescence measurement as it is separated from quencher DNP, excitation at 360 nm and absorbance measurement at 450 nm) in aqueous phase | Isopropanol | 50% | 102 | 72 | 82 | % | Incubation: 50 mM Tris-Hcl, 5mM CaCl2, Assay: 50 mM MES buffer, 10 mM CaCl2 and 0.005% Triton X-100 | Incubation: 7.5, Assay: 6.5 | Incubation: 37Β°C, Assay: 25Β°C | MOCAc-PLGL-A2pr(Dnp)-AR-NH2 | L-A2pr(Dnp)-AR-NH2 , βMOCAc-PLG | β | β | Non-incubated control (in %), assay in aqueous phase |
| N123D | Stability - Incubation | Activity after incubation (3 hours at 37Β°C), measured by fluorescence spectrophotometry (fluorophore MOCAc fluorescence measurement as it is separated from quencher DNP, excitation at 360 nm and absorbance measurement at 450 nm) in aqueous phase | Isopropanol | 50% | 92 | 38 | 66 | % | Incubation: 50 mM Tris-Hcl, 5mM CaCl2, Assay: 50 mM MES buffer, 10 mM CaCl2 and 0.005% Triton X-100 | Incubation: 7.5, Assay: 6.5 | Incubation: 37Β°C, Assay: 25Β°C | MOCAc-PLGL-A2pr(Dnp)-AR-NH2 | L-A2pr(Dnp)-AR-NH2 , βMOCAc-PLG | β | β | Non-incubated control (in %), assay in aqueous phase |
Mutations in this dataset (2) β Toshihiro Takahashi et al. (2005)
WT
N123D
Visualization : Stability β Incubation
Toshihiro Takahashi et al. (2005)
One bar per measurement. Colour = solvent, shade = solvent volume. β β β 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.