Short-chain dehydrogenase (gene NaSDR) from Novosphingobium aromaticivorans
Enzyme Description
Sequence
MTIALNNVVAVVTGAAGGIGRELVKAMKAANAIVIATDMAPSADVEGADHYLQHDVTSEAGWKAVAALAQEKYGRVDALVHNAGISIVTKFEDTPLSDFHRVNTVNVDSIIIGTQVLLPLLKEGGKARAGGASVVNFSSVGGLRGAAFNAAYCTSKAAVKMLSKCLGAEFAALGYNIRVNSVHPGGIDTPMLGSIMDKYVELGAAPSREVAQAAMEMRHPIGRMGRPAEMGGGVVYLCSDAASFVTCTEFVMDGGFSQV
Kai Wu et al. (2019)
β Efficient synthesis of an antiviral drug intermediate using an enhanced short-chain dehydrogenase in an aqueous-organic solvent system
Applied Microbiology and Biotechnology
Β· doi:10.1007/s00253-019-09781-4 β
Β· Activity + Stability - Conversion
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Database ID
UniProt: A4XEP2 β
Sequence Annotation
Explicit - Provided GenBank Accession Number
Protein Source
Recombinant, host bacterium Escherichia coli BL21 (DE3)
Experimental Data (7 measurements)
7 measurements
| Property | Assay | Solvent | Solvent Volume | Aqueous Reference | Measured Value | Units | Solution | pH | Temperature | Substrate(s) | Product(s) | Cofactor(s) | Shaking | Comments |
|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
| Activity + Stability - Conversion | Conversion measured by HPLC after 6 h of incubation at 30 Β°C and 20 minutes incubation at 90 Β°C in the presence of organic solvent | tert-Butyl Methyl Ether (MTBE) | 20% | 30.9 | 82.9 | % | 100 mM Tris-HCl buffer, 6% isopropanol | 8 | 30Β°C for 6 hours, 90Β°C for 20 minutes | 200 mM (3S)-3-(N-Boc-amino)-1-chloro-4-phenylbutan-1-one | (2R,3S)-N-tert-Butoxycarbonyl-3-amino-1-chloro-2-hydroxy-4-phenylbutane | 0.25 mM NAD+ | β | Classical aqueous control (in %) |
| Activity + Stability - Conversion | Conversion measured by HPLC after 6 h of incubation at 30 Β°C and 20 minutes incubation at 90 Β°C in the presence of organic solvent | n-Hexane | 20% | 30.9 | 97.0 | % | 100 mM Tris-HCl buffer, 6% isopropanol | 8 | 30Β°C for 6 hours, 90Β°C for 20 minutes | 200 mM (3S)-3-(N-Boc-amino)-1-chloro-4-phenylbutan-1-one | (2R,3S)-N-tert-Butoxycarbonyl-3-amino-1-chloro-2-hydroxy-4-phenylbutane | 0.25 mM NAD+ | β | Classical aqueous control (in %) |
| Activity + Stability - Conversion | Conversion measured by HPLC after 6 h of incubation at 30 Β°C and 20 minutes incubation at 90 Β°C in the presence of organic solvent | Toluene | 20% | 30.9 | 100.0 | % | 100 mM Tris-HCl buffer, 6% isopropanol | 8 | 30Β°C for 6 hours, 90Β°C for 20 minutes | 200 mM (3S)-3-(N-Boc-amino)-1-chloro-4-phenylbutan-1-one | (2R,3S)-N-tert-Butoxycarbonyl-3-amino-1-chloro-2-hydroxy-4-phenylbutane | 0.25 mM NAD+ | β | Classical aqueous control (in %) |
| Activity + Stability - Conversion | Conversion measured by HPLC after 6 h of incubation at 30 Β°C and 20 minutes incubation at 90 Β°C in the presence of organic solvent | Dimethylformamide (DMF) | 20% | 30.9 | 51.0 | % | 100 mM Tris-HCl buffer, 6% isopropanol | 8 | 30Β°C for 6 hours, 90Β°C for 20 minutes | 200 mM (3S)-3-(N-Boc-amino)-1-chloro-4-phenylbutan-1-one | (2R,3S)-N-tert-Butoxycarbonyl-3-amino-1-chloro-2-hydroxy-4-phenylbutane | 0.25 mM NAD+ | β | Classical aqueous control (in %) |
| Activity + Stability - Conversion | Conversion measured by HPLC after 6 h of incubation at 30 Β°C and 20 minutes incubation at 90 Β°C in the presence of organic solvent | Dimethyl Sulfoxide (DMSO) | 20% | 30.9 | 47.0 | % | 100 mM Tris-HCl buffer, 6% isopropanol | 8 | 30Β°C for 6 hours, 90Β°C for 20 minutes | 200 mM (3S)-3-(N-Boc-amino)-1-chloro-4-phenylbutan-1-one | (2R,3S)-N-tert-Butoxycarbonyl-3-amino-1-chloro-2-hydroxy-4-phenylbutane | 0.25 mM NAD+ | β | Classical aqueous control (in %) |
| Activity + Stability - Conversion | Conversion measured by HPLC after 6 h of incubation at 30 Β°C and 20 minutes incubation at 90 Β°C in the presence of organic solvent | Methanol | 20% | 30.9 | 19.1 | % | 100 mM Tris-HCl buffer, 6% isopropanol | 8 | 30Β°C for 6 hours, 90Β°C for 20 minutes | 200 mM (3S)-3-(N-Boc-amino)-1-chloro-4-phenylbutan-1-one | (2R,3S)-N-tert-Butoxycarbonyl-3-amino-1-chloro-2-hydroxy-4-phenylbutane | 0.25 mM NAD+ | β | Classical aqueous control (in %) |
| Activity + Stability - Conversion | Conversion measured by HPLC after 6 h of incubation at 30 Β°C and 20 minutes incubation at 90 Β°C in the presence of organic solvent | Acetonitrile | 20% | 30.9 | 13.0 | % | 100 mM Tris-HCl buffer, 6% isopropanol | 8 | 30Β°C for 6 hours, 90Β°C for 20 minutes | 200 mM (3S)-3-(N-Boc-amino)-1-chloro-4-phenylbutan-1-one | (2R,3S)-N-tert-Butoxycarbonyl-3-amino-1-chloro-2-hydroxy-4-phenylbutane | 0.25 mM NAD+ | β | Classical aqueous control (in %) |
Visualization : Activity + Stability β Conversion
One bar per measurement. Colour = solvent, shade = solvent volume.