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 2 of 2
| Property | Assay | Solvent | Solvent Volume | Aqueous Reference | Measured Value | Units | Solution | pH | Temperature | Substrate(s) | Product(s) | Cofactor(s) | Shaking | Comments |
|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
| Specificity - Enantioselectivity | Enantiomeric excess (toward the S form) after 40 minutes of incubation in the presence of organic solvent, measured by HPLC | Glycerol | 80% | 98 | 98 | % | 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 | Ethylene Glycol | 80% | 98 | 82 | % | 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 | Acetonitrile | 80% | 98 | 96 | % | 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 | Acetone | 80% | 98 | 89 | % | 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 %) |
| Stability - Incubation | Activity after incubation (24 hours at 30Β°C), measured by HPLC in aqueous phase | Toluene | 65 ppm water | 100 | 96 | % | Incubation: /, Assay: 50 mM phosphate buffer | Incubation: ?, Assay: 7.5 | Incubation: 30Β°C, Assay: 30Β°C | 50 mM Bicyclo(2.2.2)octane-2,6-dione | (S)-2-(3-ketocyclohexyl)acetic acid | β | Incubation: 120 rpm | Non-incubated control (in %), activity measured in aqueous phase | Clear about assay in aqueuos phase |
| Stability - Incubation | Activity after incubation (24 hours at 30Β°C), measured by HPLC in aqueous phase | Methanol | 26 ppm water | 100 | 4 | % | Incubation: /, Assay: 50 mM phosphate buffer | Incubation: ?, Assay: 7.5 | Incubation: 30Β°C, Assay: 30Β°C | 50 mM Bicyclo(2.2.2)octane-2,6-dione | (S)-2-(3-ketocyclohexyl)acetic acid | β | Incubation: 120 rpm | Non-incubated control (in %), activity measured in aqueous phase | Clear about assay in aqueuos phase |
| Stability - Incubation | Activity after incubation (24 hours at 30Β°C), measured by HPLC in aqueous phase | Ethanol | 14 ppm water | 100 | 58 | % | Incubation: /, Assay: 50 mM phosphate buffer | Incubation: ?, Assay: 7.5 | Incubation: 30Β°C, Assay: 30Β°C | 50 mM Bicyclo(2.2.2)octane-2,6-dione | (S)-2-(3-ketocyclohexyl)acetic acid | β | Incubation: 120 rpm | Non-incubated control (in %), activity measured in aqueous phase | Clear about assay in aqueuos phase |
| Stability - Incubation | Activity after incubation (24 hours at 30Β°C), measured by HPLC in aqueous phase | Diisopropyl Ether | 10 ppm water | 100 | 99 | % | Incubation: /, Assay: 50 mM phosphate buffer | Incubation: ?, Assay: 7.5 | Incubation: 30Β°C, Assay: 30Β°C | 50 mM Bicyclo(2.2.2)octane-2,6-dione | (S)-2-(3-ketocyclohexyl)acetic acid | β | Incubation: 120 rpm | Non-incubated control (in %), activity measured in aqueous phase | Clear about assay in aqueuos phase |
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.