S-adenosylmethionine from Acacia koa
Enzyme Description
Sequence
METFLYTSESVNEGHPDKLCDQISDAVLDACLEQDPDSKVACETCTKTNMVMVFGEITTKANVDYEKIVRNTCRNIGFVSDDVGLDADNCKVLVNIEQQSPDIAQGVHGHLTKRPEEIGAGDQGHMFGYATDETPELMPLSHVLATKLGARLTEVRKNGTCPWLRPDGKTQVTVEYYNDKGAMVPIRVHTVLISTQHDETVTNDEIAADLKEHVIKPVIPEKYLDEKTIFHLNPSGRFVIGGPHGDAGLTGRKIIIDTYGGWGAHGGGAFSGKDPTKVDRSGAYIVRQAAKSIVANGLARRCLVQVSYAIGVPEPLSVFVDSYGTGKIPDKEILKIVKENFDFRPGMISIHLDLKRGGNGRFLKTAAYGHFGREDPDFTWETVKPLKWEKPQA
James T. Carrillo et al. (2024)
β Characterization of a plant S-adenosylmethionine synthetase from Acacia koa
Plant Physiology and Biochemistry
Β· doi:10.1016/j.plaphy.2024.108618 β
Β· Activity - Michaelis-Menten Activity + Stability - Product Formation
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Database ID
UniProt: A0A9E8G339 β
Sequence Annotation
Explicit - Provided GenBank Accession Number
Protein Source
Recombinant, host bacterium Escherichia coli BL21 (DE3)
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 - Michaelis-Menten | kcat (1/s) measured by HPLC in the presence of organic solvent | Methanol | 25% | 1.69 | 1.86 | 1/s | 100 mM Tris-HCl buffer, 20 mM KCl, 10 mM MgClβ, 5 mM DTT | 8.5 | 35Β°C | ATP (Adenosine triphosphate), 4 mM L-methionine | Piβ , PPiβ , S-adenosyl-L-methionine | β | β | Classical aqueous control (in 1/s) |
| Activity - Michaelis-Menten | kcat (1/s) measured by HPLC in the presence of organic solvent | Formamide | 25% | 1.69 | 1.51 | 1/s | 100 mM Tris-HCl buffer, 20 mM KCl, 10 mM MgClβ, 5 mM DTT | 8.5 | 35Β°C | ATP (Adenosine triphosphate), 4 mM L-methionine | Piβ , PPiβ , S-adenosyl-L-methionine | β | β | Classical aqueous control (in 1/s) |
| Activity - Michaelis-Menten | kcat (1/s) measured by HPLC in the presence of organic solvent | Acetonitrile | 25% | 1.69 | 1.77 | 1/s | 100 mM Tris-HCl buffer, 20 mM KCl, 10 mM MgClβ, 5 mM DTT | 8.5 | 35Β°C | ATP (Adenosine triphosphate), 4 mM L-methionine | Piβ , PPiβ , S-adenosyl-L-methionine | β | β | Classical aqueous control (in 1/s) |
| Activity + Stability - Product Formation | Relative product formation after 1 hour incubation at 35Β°C, measured by HPLC in the presence of organic solvent | Dimethyl Sulfoxide (DMSO) | 50% | 1.0 | 0.12 | relative units, 1 for control | 100 mM Tris-HCl buffer, 20 mM KCl, 10 mM MgClβ, 5 mM DTT | 8.5 | 35Β°C | ATP (Adenosine triphosphate), 4 mM L-methionine | Piβ , PPiβ , S-adenosyl-L-methionine | β | β | Classical aqueous control (in relative units, 1 for control) |
| Activity + Stability - Product Formation | Relative product formation after 1 hour incubation at 35Β°C, measured by HPLC in the presence of organic solvent | Dimethyl Sulfoxide (DMSO) | 25% | 1.0 | 0.54 | relative units, 1 for control | 100 mM Tris-HCl buffer, 20 mM KCl, 10 mM MgClβ, 5 mM DTT | 8.5 | 35Β°C | ATP (Adenosine triphosphate), 4 mM L-methionine | Piβ , PPiβ , S-adenosyl-L-methionine | β | β | Classical aqueous control (in relative units, 1 for control) |
| Activity + Stability - Product Formation | Relative product formation after 1 hour incubation at 35Β°C, measured by HPLC in the presence of organic solvent | Dimethyl Sulfoxide (DMSO) | 10% | 1.0 | 0.49 | relative units, 1 for control | 100 mM Tris-HCl buffer, 20 mM KCl, 10 mM MgClβ, 5 mM DTT | 8.5 | 35Β°C | ATP (Adenosine triphosphate), 4 mM L-methionine | Piβ , PPiβ , S-adenosyl-L-methionine | β | β | Classical aqueous control (in relative units, 1 for control) |
| Activity + Stability - Product Formation | Relative product formation after 1 hour incubation at 35Β°C, measured by HPLC in the presence of organic solvent | Dimethyl Sulfoxide (DMSO) | 5% | 1.0 | 0.58 | relative units, 1 for control | 100 mM Tris-HCl buffer, 20 mM KCl, 10 mM MgClβ, 5 mM DTT | 8.5 | 35Β°C | ATP (Adenosine triphosphate), 4 mM L-methionine | Piβ , PPiβ , S-adenosyl-L-methionine | β | β | Classical aqueous control (in relative units, 1 for control) |
| Activity + Stability - Product Formation | Relative product formation after 1 hour incubation at 35Β°C, measured by HPLC in the presence of organic solvent | Formamide | 50% | 1.0 | 0.24 | relative units, 1 for control | 100 mM Tris-HCl buffer, 20 mM KCl, 10 mM MgClβ, 5 mM DTT | 8.5 | 35Β°C | ATP (Adenosine triphosphate), 4 mM L-methionine | Piβ , PPiβ , S-adenosyl-L-methionine | β | β | Classical aqueous control (in relative units, 1 for control) |
| Activity + Stability - Product Formation | Relative product formation after 1 hour incubation at 35Β°C, measured by HPLC in the presence of organic solvent | Formamide | 25% | 1.0 | 1.36 | relative units, 1 for control | 100 mM Tris-HCl buffer, 20 mM KCl, 10 mM MgClβ, 5 mM DTT | 8.5 | 35Β°C | ATP (Adenosine triphosphate), 4 mM L-methionine | Piβ , PPiβ , S-adenosyl-L-methionine | β | β | Classical aqueous control (in relative units, 1 for control) |
| Activity + Stability - Product Formation | Relative product formation after 1 hour incubation at 35Β°C, measured by HPLC in the presence of organic solvent | Formamide | 10% | 1.0 | 0.76 | relative units, 1 for control | 100 mM Tris-HCl buffer, 20 mM KCl, 10 mM MgClβ, 5 mM DTT | 8.5 | 35Β°C | ATP (Adenosine triphosphate), 4 mM L-methionine | Piβ , PPiβ , S-adenosyl-L-methionine | β | β | Classical aqueous control (in relative units, 1 for control) |
| Activity + Stability - Product Formation | Relative product formation after 1 hour incubation at 35Β°C, measured by HPLC in the presence of organic solvent | Formamide | 5% | 1.0 | 0.58 | relative units, 1 for control | 100 mM Tris-HCl buffer, 20 mM KCl, 10 mM MgClβ, 5 mM DTT | 8.5 | 35Β°C | ATP (Adenosine triphosphate), 4 mM L-methionine | Piβ , PPiβ , S-adenosyl-L-methionine | β | β | Classical aqueous control (in relative units, 1 for control) |
| Activity + Stability - Product Formation | Relative product formation after 1 hour incubation at 35Β°C, measured by HPLC in the presence of organic solvent | Formamide | 50% | 1.0 | 0.1 | relative units, 1 for control | 100 mM Tris-HCl buffer, 20 mM KCl, 10 mM MgClβ, 5 mM DTT | 8.5 | 35Β°C | ATP (Adenosine triphosphate), 4 mM L-methionine | Piβ , PPiβ , S-adenosyl-L-methionine | β | β | Classical aqueous control (in relative units, 1 for control) |
| Activity + Stability - Product Formation | Relative product formation after 1 hour incubation at 35Β°C, measured by HPLC in the presence of organic solvent | Formamide | 25% | 1.0 | 0.24 | relative units, 1 for control | 100 mM Tris-HCl buffer, 20 mM KCl, 10 mM MgClβ, 5 mM DTT | 8.5 | 35Β°C | ATP (Adenosine triphosphate), 4 mM L-methionine | Piβ , PPiβ , S-adenosyl-L-methionine | β | β | Classical aqueous control (in relative units, 1 for control) |
| Activity + Stability - Product Formation | Relative product formation after 1 hour incubation at 35Β°C, measured by HPLC in the presence of organic solvent | Formamide | 10% | 1.0 | 0.37 | relative units, 1 for control | 100 mM Tris-HCl buffer, 20 mM KCl, 10 mM MgClβ, 5 mM DTT | 8.5 | 35Β°C | ATP (Adenosine triphosphate), 4 mM L-methionine | Piβ , PPiβ , S-adenosyl-L-methionine | β | β | Classical aqueous control (in relative units, 1 for control) |
| Activity + Stability - Product Formation | Relative product formation after 1 hour incubation at 35Β°C, measured by HPLC in the presence of organic solvent | Formamide | 5% | 1.0 | 0.56 | relative units, 1 for control | 100 mM Tris-HCl buffer, 20 mM KCl, 10 mM MgClβ, 5 mM DTT | 8.5 | 35Β°C | ATP (Adenosine triphosphate), 4 mM L-methionine | Piβ , PPiβ , S-adenosyl-L-methionine | β | β | Classical aqueous control (in relative units, 1 for control) |
| Activity + Stability - Product Formation | Relative product formation after 1 hour incubation at 35Β°C, measured by HPLC in the presence of organic solvent | Acetonitrile | 25% | 1.0 | 1.3 | relative units, 1 for control | 100 mM Tris-HCl buffer, 20 mM KCl, 10 mM MgClβ, 5 mM DTT | 8.5 | 35Β°C | ATP (Adenosine triphosphate), 4 mM L-methionine | Piβ , PPiβ , S-adenosyl-L-methionine | β | β | Classical aqueous control (in relative units, 1 for control) |
| Activity + Stability - Product Formation | Relative product formation after 1 hour incubation at 35Β°C, measured by HPLC in the presence of organic solvent | Acetonitrile | 10% | 1.0 | 0.8 | relative units, 1 for control | 100 mM Tris-HCl buffer, 20 mM KCl, 10 mM MgClβ, 5 mM DTT | 8.5 | 35Β°C | ATP (Adenosine triphosphate), 4 mM L-methionine | Piβ , PPiβ , S-adenosyl-L-methionine | β | β | Classical aqueous control (in relative units, 1 for control) |
| Activity + Stability - Product Formation | Relative product formation after 1 hour incubation at 35Β°C, measured by HPLC in the presence of organic solvent | Acetonitrile | 5% | 1.0 | 0.6 | relative units, 1 for control | 100 mM Tris-HCl buffer, 20 mM KCl, 10 mM MgClβ, 5 mM DTT | 8.5 | 35Β°C | ATP (Adenosine triphosphate), 4 mM L-methionine | Piβ , PPiβ , S-adenosyl-L-methionine | β | β | Classical aqueous control (in relative units, 1 for control) |
| Activity + Stability - Product Formation | Relative product formation after 1 hour incubation at 35Β°C, measured by HPLC in the presence of organic solvent | Methanol | 50% | 1.0 | 0.7 | relative units, 1 for control | 100 mM Tris-HCl buffer, 20 mM KCl, 10 mM MgClβ, 5 mM DTT | 8.5 | 35Β°C | ATP (Adenosine triphosphate), 4 mM L-methionine | Piβ , PPiβ , S-adenosyl-L-methionine | β | β | Classical aqueous control (in relative units, 1 for control) |
| Activity + Stability - Product Formation | Relative product formation after 1 hour incubation at 35Β°C, measured by HPLC in the presence of organic solvent | Methanol | 25% | 1.0 | 1.6 | relative units, 1 for control | 100 mM Tris-HCl buffer, 20 mM KCl, 10 mM MgClβ, 5 mM DTT | 8.5 | 35Β°C | ATP (Adenosine triphosphate), 4 mM L-methionine | Piβ , PPiβ , S-adenosyl-L-methionine | β | β | Classical aqueous control (in relative units, 1 for control) |
Visualization : Activity β Michaelis-Menten
One bar per measurement. Colour = solvent, shade = solvent volume.
Visualization : Activity + Stability β Product Formation
One bar per measurement. Colour = solvent, shade = solvent volume.