6-oxo camphor hydrolase (gene camK) from Rhodococcus sp.
Enzyme Description
Sequence
MKQLATPFQEYSQKYENIRLERDGGVLLVTVHTEGKSLVWTSTAHDELAYCFHDIACDRENKVVILTGTGPSFCNEIDFTSFNLGTPHDWDEIIFEGQRLLNNLLSIEVPVIAAVNGPVTNHPEIPVMSDIVLAAESATFQDGPHFPSGIVPGDGAHVVWPHVLGSNRGRYFLLTGQELDARTALDYGAVNEVLSEQELLPRAWELARGIAEKPLLARRYARKVLTRQLRRVMEADLSLGLAHEALAAIDLGMESEQ
Elina Siirola et al. (2011)
β Tolerance of Ξ²-diketone hydrolases as representatives of the crotonase superfamily towards organic solvents
Biotechnology and Bioengineering
Β· doi:10.1002/bit.23275 β
Β· Activity - Classical Stability - Incubation Specificity - Enantioselectivity
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Database ID
UniProt: Q93TU6 β
Sequence Annotation
Inferred - from protein name
Protein Source
Recombinant, host bacterium Escherichia coli BL21 (DE3)
Experimental Data (28 measurements)
28 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 HPLC in the presence of organic solvent | Petroleum Ether | 80% | 100 | 49 | % | 50 mM phosphate buffer | 7.5 | 30Β°C | 50 mM Bicyclo(2.2.2)octane-2,6-dione | (S)-2-(3-ketocyclohexyl)acetic acid | β | 120 rpm | Classical aqueous control (in %) |
| Activity - Classical | Activity measured by HPLC in the presence of organic solvent | Diisopropyl Ether | 80% | 100 | 51 | % | 50 mM phosphate buffer | 7.5 | 30Β°C | 50 mM Bicyclo(2.2.2)octane-2,6-dione | (S)-2-(3-ketocyclohexyl)acetic acid | β | 120 rpm | Classical aqueous control (in %) |
| Activity - Classical | Activity measured by HPLC in the presence of organic solvent | tert-Butyl Methyl Ether (MTBE) | 80% | 100 | 5 | % | 50 mM phosphate buffer | 7.5 | 30Β°C | 50 mM Bicyclo(2.2.2)octane-2,6-dione | (S)-2-(3-ketocyclohexyl)acetic acid | β | 120 rpm | Classical aqueous control (in %) |
| Activity - Classical | Activity measured by HPLC in the presence of organic solvent | n-Hexane | 80% | 100 | 51 | % | 50 mM phosphate buffer | 7.5 | 30Β°C | 50 mM Bicyclo(2.2.2)octane-2,6-dione | (S)-2-(3-ketocyclohexyl)acetic acid | β | 120 rpm | Classical aqueous control (in %) |
| Activity - Classical | Activity measured by HPLC in the presence of organic solvent | Diethyl Ether | 80% | 100 | 32 | % | 50 mM phosphate buffer | 7.5 | 30Β°C | 50 mM Bicyclo(2.2.2)octane-2,6-dione | (S)-2-(3-ketocyclohexyl)acetic acid | β | 120 rpm | Classical aqueous control (in %) |
| Activity - Classical | Activity measured by HPLC in the presence of organic solvent | Toluene | 80% | 100 | 29 | % | 50 mM phosphate buffer | 7.5 | 30Β°C | 50 mM Bicyclo(2.2.2)octane-2,6-dione | (S)-2-(3-ketocyclohexyl)acetic acid | β | 120 rpm | Classical aqueous control (in %) |
| Activity - Classical | Activity measured by HPLC in the presence of organic solvent | Ethyl Acetate | 80% | 100 | 12 | % | 50 mM phosphate buffer | 7.5 | 30Β°C | 50 mM Bicyclo(2.2.2)octane-2,6-dione | (S)-2-(3-ketocyclohexyl)acetic acid | β | 120 rpm | Classical aqueous control (in %) |
| Activity - Classical | Activity measured by HPLC in the presence of organic solvent | Tetrahydrofuran (THF) | 80% | 100 | 0 | % | 50 mM phosphate buffer | 7.5 | 30Β°C | 50 mM Bicyclo(2.2.2)octane-2,6-dione | (S)-2-(3-ketocyclohexyl)acetic acid | β | 120 rpm | Classical aqueous control (in %) |
| Activity - Classical | Activity measured by HPLC in the presence of organic solvent | Glycerol | 80% | 100 | 5 | % | 50 mM phosphate buffer | 7.5 | 30Β°C | 50 mM Bicyclo(2.2.2)octane-2,6-dione | (S)-2-(3-ketocyclohexyl)acetic acid | β | 120 rpm | Classical aqueous control (in %) |
| Activity - Classical | Activity measured by HPLC in the presence of organic solvent | Ethylene Glycol | 80% | 100 | 5 | % | 50 mM phosphate buffer | 7.5 | 30Β°C | 50 mM Bicyclo(2.2.2)octane-2,6-dione | (S)-2-(3-ketocyclohexyl)acetic acid | β | 120 rpm | Classical aqueous control (in %) |
| Activity - Classical | Activity measured by HPLC in the presence of organic solvent | Acetonitrile | 80% | 100 | 5 | % | 50 mM phosphate buffer | 7.5 | 30Β°C | 50 mM Bicyclo(2.2.2)octane-2,6-dione | (S)-2-(3-ketocyclohexyl)acetic acid | β | 120 rpm | Classical aqueous control (in %) |
| Activity - Classical | Activity measured by HPLC in the presence of organic solvent | 1,4-Dioxane | 80% | 100 | 0 | % | 50 mM phosphate buffer | 7.5 | 30Β°C | 50 mM Bicyclo(2.2.2)octane-2,6-dione | (S)-2-(3-ketocyclohexyl)acetic acid | β | 120 rpm | Classical aqueous control (in %) |
| Activity - Classical | Activity measured by HPLC in the presence of organic solvent | Acetone | 80% | 100 | 5 | % | 50 mM phosphate buffer | 7.5 | 30Β°C | 50 mM Bicyclo(2.2.2)octane-2,6-dione | (S)-2-(3-ketocyclohexyl)acetic acid | β | 120 rpm | Classical aqueous control (in %) |
| Specificity - Enantioselectivity | Enantiomeric excess (toward the S form) after 40 minutes of incubation in the presence of organic solvent, measured by HPLC | Petroleum Ether | 80% | 98 | >99 | % | 50 mM phosphate buffer | 7.5 | 30Β°C | 50 mM Bicyclo(2.2.2)octane-2,6-dione | (S)-2-(3-ketocyclohexyl)acetic acid | β | 120 rpm | Classical aqueous control (in %) |
| Specificity - Enantioselectivity | Enantiomeric excess (toward the S form) after 40 minutes of incubation in the presence of organic solvent, measured by HPLC | Diisopropyl Ether | 80% | 98 | 99 | % | 50 mM phosphate buffer | 7.5 | 30Β°C | 50 mM Bicyclo(2.2.2)octane-2,6-dione | (S)-2-(3-ketocyclohexyl)acetic acid | β | 120 rpm | Classical aqueous control (in %) |
| Specificity - Enantioselectivity | Enantiomeric excess (toward the S form) after 40 minutes of incubation in the presence of organic solvent, measured by HPLC | tert-Butyl Methyl Ether (MTBE) | 80% | 98 | 93 | % | 50 mM phosphate buffer | 7.5 | 30Β°C | 50 mM Bicyclo(2.2.2)octane-2,6-dione | (S)-2-(3-ketocyclohexyl)acetic acid | β | 120 rpm | Classical aqueous control (in %) |
| Specificity - Enantioselectivity | Enantiomeric excess (toward the S form) after 40 minutes of incubation in the presence of organic solvent, measured by HPLC | n-Hexane | 80% | 98 | >99 | % | 50 mM phosphate buffer | 7.5 | 30Β°C | 50 mM Bicyclo(2.2.2)octane-2,6-dione | (S)-2-(3-ketocyclohexyl)acetic acid | β | 120 rpm | Classical aqueous control (in %) |
| Specificity - Enantioselectivity | Enantiomeric excess (toward the S form) after 40 minutes of incubation in the presence of organic solvent, measured by HPLC | Diethyl Ether | 80% | 98 | 99 | % | 50 mM phosphate buffer | 7.5 | 30Β°C | 50 mM Bicyclo(2.2.2)octane-2,6-dione | (S)-2-(3-ketocyclohexyl)acetic acid | β | 120 rpm | Classical aqueous control (in %) |
| Specificity - Enantioselectivity | Enantiomeric excess (toward the S form) after 40 minutes of incubation in the presence of organic solvent, measured by HPLC | Toluene | 80% | 98 | >99 | % | 50 mM phosphate buffer | 7.5 | 30Β°C | 50 mM Bicyclo(2.2.2)octane-2,6-dione | (S)-2-(3-ketocyclohexyl)acetic acid | β | 120 rpm | Classical aqueous control (in %) |
| Specificity - Enantioselectivity | Enantiomeric excess (toward the S form) after 40 minutes of incubation in the presence of organic solvent, measured by HPLC | Ethyl Acetate | 80% | 98 | >99 | % | 50 mM phosphate buffer | 7.5 | 30Β°C | 50 mM Bicyclo(2.2.2)octane-2,6-dione | (S)-2-(3-ketocyclohexyl)acetic acid | β | 120 rpm | Classical aqueous control (in %) |
Visualization : Activity β Classical
One bar per measurement. Colour = solvent, shade = solvent volume.
Visualization : Stability β Incubation
One bar per measurement. Colour = solvent, shade = solvent volume.
Visualization : Specificity β Enantioselectivity
One bar per measurement. Colour = solvent, shade = solvent volume.