Sortase A (gene SrtA) from Staphylococcus aureus
Enzyme Description
Sequence
MKKWTNRLMTIAGVVLILVAAYLFAKPHIDNYLHDKDKDEKIEQYDKNVKEQASKDKKQQAKPQIPKDKSKVAGYIEIPDADIKEPVYPGPATPEQLNRGVSFAEENESLDDQNISIAGHTFIDRPNYQFTNLKAAKKGSMVYFKVGNETRKYKMTSIRDVKPTDVGVLDEQKGKDKQLTLITCDDYNEKTGVWEKRKIFVATEVK
Zhimeng Wu et al. (2017)
β Efficient expression of sortase A from Staphylococcus aureus in Escherichia coli and its enzymatic characterizations
Bioresources and Bioprocessing
Β· doi:10.1186/s40643-017-0143-y β
Β· Activity - Classical
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Database ID
UniProt: Q2FV99 β
Sequence Annotation
Inferred - from protein name
Protein Source
Recombinant, host bacterium Escherichia coli BL21 (DE3)
Experimental Data (25 measurements)
25 measurements β page 1 of 2
| Property | Assay | Solvent | Solvent Volume | Aqueous Reference | Measured Value | Units | Extraction method | Data type | Solution | pH | Temperature | Substrate(s) | Product(s) | Cofactor(s) | Shaking | Comments |
|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
| Activity - Classical | Activity measured by fluorescence spectrophotometry (cleavage separates dabcyl quencher from edans acceptor, thus removing the quencher and increasing fluoresccence, 350 nm excitation, 495 nm measurement) in the presence of organic solvent | Acetonitrile | 30 % v/v | 100 | 0 | % | Digital | Continuous | 50 mM TrisβHCl buffer, 150 mM NaCl, 30 mM CaCl2 | 7.8 | 37Β°C | 0.5 Mg Dabcyl-QALPETGEE-Edans | Dabcyl-QALPET , GEE-Edans | β | β | Classical aqueous control (in %) |
| Activity - Classical | Activity measured by fluorescence spectrophotometry (cleavage separates dabcyl quencher from edans acceptor, thus removing the quencher and increasing fluoresccence, 350 nm excitation, 495 nm measurement) in the presence of organic solvent | Dimethyl Sulfoxide (DMSO) | 50 % v/v | 100 | 10 | % | Digital | Continuous | 50 mM TrisβHCl buffer, 150 mM NaCl, 30 mM CaCl2 | 7.8 | 37Β°C | 0.5 Mg Dabcyl-QALPETGEE-Edans | Dabcyl-QALPET , GEE-Edans | β | β | Classical aqueous control (in %) |
| Activity - Classical | Activity measured by fluorescence spectrophotometry (cleavage separates dabcyl quencher from edans acceptor, thus removing the quencher and increasing fluoresccence, 350 nm excitation, 495 nm measurement) in the presence of organic solvent | Dimethyl Sulfoxide (DMSO) | 40 % v/v | 100 | 70 | % | Digital | Continuous | 50 mM TrisβHCl buffer, 150 mM NaCl, 30 mM CaCl2 | 7.8 | 37Β°C | 0.5 Mg Dabcyl-QALPETGEE-Edans | Dabcyl-QALPET , GEE-Edans | β | β | Classical aqueous control (in %) |
| Activity - Classical | Activity measured by fluorescence spectrophotometry (cleavage separates dabcyl quencher from edans acceptor, thus removing the quencher and increasing fluoresccence, 350 nm excitation, 495 nm measurement) in the presence of organic solvent | Dimethyl Sulfoxide (DMSO) | 30 % v/v | 100 | 96 | % | Digital | Continuous | 50 mM TrisβHCl buffer, 150 mM NaCl, 30 mM CaCl2 | 7.8 | 37Β°C | 0.5 Mg Dabcyl-QALPETGEE-Edans | Dabcyl-QALPET , GEE-Edans | β | β | Classical aqueous control (in %) |
| Activity - Classical | Activity measured by fluorescence spectrophotometry (cleavage separates dabcyl quencher from edans acceptor, thus removing the quencher and increasing fluoresccence, 350 nm excitation, 495 nm measurement) in the presence of organic solvent | Dimethyl Sulfoxide (DMSO) | 20 % v/v | 100 | 98 | % | Digital | Continuous | 50 mM TrisβHCl buffer, 150 mM NaCl, 30 mM CaCl2 | 7.8 | 37Β°C | 0.5 Mg Dabcyl-QALPETGEE-Edans | Dabcyl-QALPET , GEE-Edans | β | β | Classical aqueous control (in %) |
| Activity - Classical | Activity measured by fluorescence spectrophotometry (cleavage separates dabcyl quencher from edans acceptor, thus removing the quencher and increasing fluoresccence, 350 nm excitation, 495 nm measurement) in the presence of organic solvent | Dimethyl Sulfoxide (DMSO) | 10 % v/v | 100 | 98 | % | Digital | Continuous | 50 mM TrisβHCl buffer, 150 mM NaCl, 30 mM CaCl2 | 7.8 | 37Β°C | 0.5 Mg Dabcyl-QALPETGEE-Edans | Dabcyl-QALPET , GEE-Edans | β | β | Classical aqueous control (in %) |
| Activity - Classical | Activity measured by fluorescence spectrophotometry (cleavage separates dabcyl quencher from edans acceptor, thus removing the quencher and increasing fluoresccence, 350 nm excitation, 495 nm measurement) in the presence of organic solvent | Acetone | 50 % v/v | 100 | 0 | % | Digital | Continuous | 50 mM TrisβHCl buffer, 150 mM NaCl, 30 mM CaCl2 | 7.8 | 37Β°C | 0.5 Mg Dabcyl-QALPETGEE-Edans | Dabcyl-QALPET , GEE-Edans | β | β | Classical aqueous control (in %) |
| Activity - Classical | Activity measured by fluorescence spectrophotometry (cleavage separates dabcyl quencher from edans acceptor, thus removing the quencher and increasing fluoresccence, 350 nm excitation, 495 nm measurement) in the presence of organic solvent | Acetone | 40 % v/v | 100 | 2 | % | Digital | Continuous | 50 mM TrisβHCl buffer, 150 mM NaCl, 30 mM CaCl2 | 7.8 | 37Β°C | 0.5 Mg Dabcyl-QALPETGEE-Edans | Dabcyl-QALPET , GEE-Edans | β | β | Classical aqueous control (in %) |
| Activity - Classical | Activity measured by fluorescence spectrophotometry (cleavage separates dabcyl quencher from edans acceptor, thus removing the quencher and increasing fluoresccence, 350 nm excitation, 495 nm measurement) in the presence of organic solvent | Acetone | 30 % v/v | 100 | 25 | % | Digital | Continuous | 50 mM TrisβHCl buffer, 150 mM NaCl, 30 mM CaCl2 | 7.8 | 37Β°C | 0.5 Mg Dabcyl-QALPETGEE-Edans | Dabcyl-QALPET , GEE-Edans | β | β | Classical aqueous control (in %) |
| Activity - Classical | Activity measured by fluorescence spectrophotometry (cleavage separates dabcyl quencher from edans acceptor, thus removing the quencher and increasing fluoresccence, 350 nm excitation, 495 nm measurement) in the presence of organic solvent | Acetone | 20 % v/v | 100 | 50 | % | Digital | Continuous | 50 mM TrisβHCl buffer, 150 mM NaCl, 30 mM CaCl2 | 7.8 | 37Β°C | 0.5 Mg Dabcyl-QALPETGEE-Edans | Dabcyl-QALPET , GEE-Edans | β | β | Classical aqueous control (in %) |
| Activity - Classical | Activity measured by fluorescence spectrophotometry (cleavage separates dabcyl quencher from edans acceptor, thus removing the quencher and increasing fluoresccence, 350 nm excitation, 495 nm measurement) in the presence of organic solvent | Acetone | 10 % v/v | 100 | 74 | % | Digital | Continuous | 50 mM TrisβHCl buffer, 150 mM NaCl, 30 mM CaCl2 | 7.8 | 37Β°C | 0.5 Mg Dabcyl-QALPETGEE-Edans | Dabcyl-QALPET , GEE-Edans | β | β | Classical aqueous control (in %) |
| Activity - Classical | Activity measured by fluorescence spectrophotometry (cleavage separates dabcyl quencher from edans acceptor, thus removing the quencher and increasing fluoresccence, 350 nm excitation, 495 nm measurement) in the presence of organic solvent | Acetonitrile | 50 % v/v | 100 | 0 | % | Digital | Continuous | 50 mM TrisβHCl buffer, 150 mM NaCl, 30 mM CaCl2 | 7.8 | 37Β°C | 0.5 Mg Dabcyl-QALPETGEE-Edans | Dabcyl-QALPET , GEE-Edans | β | β | Classical aqueous control (in %) |
| Activity - Classical | Activity measured by fluorescence spectrophotometry (cleavage separates dabcyl quencher from edans acceptor, thus removing the quencher and increasing fluoresccence, 350 nm excitation, 495 nm measurement) in the presence of organic solvent | Acetonitrile | 40 % v/v | 100 | 0 | % | Digital | Continuous | 50 mM TrisβHCl buffer, 150 mM NaCl, 30 mM CaCl2 | 7.8 | 37Β°C | 0.5 Mg Dabcyl-QALPETGEE-Edans | Dabcyl-QALPET , GEE-Edans | β | β | Classical aqueous control (in %) |
| Activity - Classical | Activity measured by fluorescence spectrophotometry (cleavage separates dabcyl quencher from edans acceptor, thus removing the quencher and increasing fluoresccence, 350 nm excitation, 495 nm measurement) in the presence of organic solvent | Methanol | 10 % v/v | 100 | 73 | % | Digital | Continuous | 50 mM TrisβHCl buffer, 150 mM NaCl, 30 mM CaCl2 | 7.8 | 37Β°C | 0.5 Mg Dabcyl-QALPETGEE-Edans | Dabcyl-QALPET , GEE-Edans | β | β | Classical aqueous control (in %) |
| Activity - Classical | Activity measured by fluorescence spectrophotometry (cleavage separates dabcyl quencher from edans acceptor, thus removing the quencher and increasing fluoresccence, 350 nm excitation, 495 nm measurement) in the presence of organic solvent | Acetonitrile | 20 % v/v | 100 | 26 | % | Digital | Continuous | 50 mM TrisβHCl buffer, 150 mM NaCl, 30 mM CaCl2 | 7.8 | 37Β°C | 0.5 Mg Dabcyl-QALPETGEE-Edans | Dabcyl-QALPET , GEE-Edans | β | β | Classical aqueous control (in %) |
| Activity - Classical | Activity measured by fluorescence spectrophotometry (cleavage separates dabcyl quencher from edans acceptor, thus removing the quencher and increasing fluoresccence, 350 nm excitation, 495 nm measurement) in the presence of organic solvent | Acetonitrile | 10 % v/v | 100 | 68 | % | Digital | Continuous | 50 mM TrisβHCl buffer, 150 mM NaCl, 30 mM CaCl2 | 7.8 | 37Β°C | 0.5 Mg Dabcyl-QALPETGEE-Edans | Dabcyl-QALPET , GEE-Edans | β | β | Classical aqueous control (in %) |
| Activity - Classical | Activity measured by fluorescence spectrophotometry (cleavage separates dabcyl quencher from edans acceptor, thus removing the quencher and increasing fluoresccence, 350 nm excitation, 495 nm measurement) in the presence of organic solvent | Ethanol | 50 % v/v | 100 | 0 | % | Digital | Continuous | 50 mM TrisβHCl buffer, 150 mM NaCl, 30 mM CaCl2 | 7.8 | 37Β°C | 0.5 Mg Dabcyl-QALPETGEE-Edans | Dabcyl-QALPET , GEE-Edans | β | β | Classical aqueous control (in %) |
| Activity - Classical | Activity measured by fluorescence spectrophotometry (cleavage separates dabcyl quencher from edans acceptor, thus removing the quencher and increasing fluoresccence, 350 nm excitation, 495 nm measurement) in the presence of organic solvent | Ethanol | 40 % v/v | 100 | 0 | % | Digital | Continuous | 50 mM TrisβHCl buffer, 150 mM NaCl, 30 mM CaCl2 | 7.8 | 37Β°C | 0.5 Mg Dabcyl-QALPETGEE-Edans | Dabcyl-QALPET , GEE-Edans | β | β | Classical aqueous control (in %) |
| Activity - Classical | Activity measured by fluorescence spectrophotometry (cleavage separates dabcyl quencher from edans acceptor, thus removing the quencher and increasing fluoresccence, 350 nm excitation, 495 nm measurement) in the presence of organic solvent | Ethanol | 30 % v/v | 100 | 1 | % | Digital | Continuous | 50 mM TrisβHCl buffer, 150 mM NaCl, 30 mM CaCl2 | 7.8 | 37Β°C | 0.5 Mg Dabcyl-QALPETGEE-Edans | Dabcyl-QALPET , GEE-Edans | β | β | Classical aqueous control (in %) |
| Activity - Classical | Activity measured by fluorescence spectrophotometry (cleavage separates dabcyl quencher from edans acceptor, thus removing the quencher and increasing fluoresccence, 350 nm excitation, 495 nm measurement) in the presence of organic solvent | Ethanol | 20 % v/v | 100 | 17 | % | Digital | Continuous | 50 mM TrisβHCl buffer, 150 mM NaCl, 30 mM CaCl2 | 7.8 | 37Β°C | 0.5 Mg Dabcyl-QALPETGEE-Edans | Dabcyl-QALPET , GEE-Edans | β | β | Classical aqueous control (in %) |
Visualization : Activity β Classical
One bar per measurement. Colour = solvent, shade = solvent volume.