Ene-reductase NCR from Zymomonas mobilis

Enzyme Description

Extremophile
No
EC Number

Sequence

Length: 358 amino acids
MPSLFDPIRFGAFTAKNRIWMAPLTRGRATRDHVPTEIMAEYYAQRASAGLIISEATGISQEGLGWPYAPGIWSDAQVEAWLPITQAVHDAGGLIFAQLWHMGRMVPSNVSGMQPVAPSASQAPGLGHTYDGKKPYDVARALRLDEIPRLLDDYEKAARHALKAGFDGVQIHAANGYLIDEFIRDSTNHRHDEYGGAVENRIRLLKDVTERVIATIGKERTAVRLSPNGEIQGTVDSHPEQVFIPAAKMLSDLDIAFLGMREGAVDGTFGKTDQPKLSPEIRKVFKPPLVLNQDYTFETAQAALDSGVADAISFGRPFIGNPDLPRRFFEKAPLTKDVIETWYTQTPKGYTDYPLLGD
Sabrina Reich et al. (2014) β€” Variations in the stability of NCR ene reductase by rational enzyme loop modulation
Journal of Structural Biology  Β· doi:10.1016/j.jsb.2013.04.004 β†—  Β· Activity + Stability - Product Formation
48 measurements
Database ID
UniProt: Q5NLA1 β†—
Sequence Annotation
Explicit - Provided
Protein Source
Recombinant, host bacterium Escherichia coli BL21 (DE3)

Experimental Data (48 measurements)

48 measurements β€” page 2 of 3
Mutation Property Assay Solvent Solvent Volume Aqueous ReferenceWT Reference Measured Value Units Solution pH Temperature Substrate(s) Product(s) Cofactor(s) Shaking Comments
DEL267-270 Activity + Stability - Product Formation Product yield after incubaton (4 hours at 30Β°C) in the presence of organic solvent, measured by HPLC Isopropanol 5% 99.6 83.6 81.3 % 50 mM Tris-HCl 7.5 30Β°C 4 mM Ketoisophorone Levodione 4 mM NADH 180 rpm Classical aqueous control (in %)
DEL267-270 Activity + Stability - Product Formation Product yield after incubaton (4 hours at 30Β°C) in the presence of organic solvent, measured by HPLC Isopropanol 10% 99.6 64.0 85.3 % 50 mM Tris-HCl 7.5 30Β°C 4 mM Ketoisophorone Levodione 4 mM NADH 180 rpm Classical aqueous control (in %)
DEL267-270 Activity + Stability - Product Formation Product yield after incubaton (4 hours at 30Β°C) in the presence of organic solvent, measured by HPLC Isopropanol 20% 99.6 60.4 79.6 % 50 mM Tris-HCl 7.5 30Β°C 4 mM Ketoisophorone Levodione 4 mM NADH 180 rpm Classical aqueous control (in %)
DEL267-270 Activity + Stability - Product Formation Product yield after incubaton (4 hours at 30Β°C) in the presence of organic solvent, measured by HPLC Isopropanol 30% 99.6 33.8 68.0 % 50 mM Tris-HCl 7.5 30Β°C 4 mM Ketoisophorone Levodione 4 mM NADH 180 rpm Classical aqueous control (in %)
DEL267-270 Activity + Stability - Product Formation Product yield after incubaton (4 hours at 30Β°C) in the presence of organic solvent, measured by HPLC Ethyl Acetate 5% 99.6 79.8 80.3 % 50 mM Tris-HCl 7.5 30Β°C 4 mM Ketoisophorone Levodione 4 mM NADH 180 rpm Classical aqueous control (in %)
DEL267-270 Activity + Stability - Product Formation Product yield after incubaton (4 hours at 30Β°C) in the presence of organic solvent, measured by HPLC Ethyl Acetate 10% 99.6 65.0 87.4 % 50 mM Tris-HCl 7.5 30Β°C 4 mM Ketoisophorone Levodione 4 mM NADH 180 rpm Classical aqueous control (in %)
DEL267-270 Activity + Stability - Product Formation Product yield after incubaton (4 hours at 30Β°C) in the presence of organic solvent, measured by HPLC Ethyl Acetate 20% 99.6 36.3 74.4 % 50 mM Tris-HCl 7.5 30Β°C 4 mM Ketoisophorone Levodione 4 mM NADH 180 rpm Classical aqueous control (in %)
DEL267-270 Activity + Stability - Product Formation Product yield after incubaton (4 hours at 30Β°C) in the presence of organic solvent, measured by HPLC Ethyl Acetate 30% 99.6 23.8 57.4 % 50 mM Tris-HCl 7.5 30Β°C 4 mM Ketoisophorone Levodione 4 mM NADH 180 rpm Classical aqueous control (in %)
DEL267-270 Activity + Stability - Product Formation Product yield after incubaton (4 hours at 30Β°C) in the presence of organic solvent, measured by HPLC Tetrahydrofuran (THF) 5% 99.6 90.2 98.2 % 50 mM Tris-HCl 7.5 30Β°C 4 mM Ketoisophorone Levodione 4 mM NADH 180 rpm Classical aqueous control (in %)
DEL267-270 Activity + Stability - Product Formation Product yield after incubaton (4 hours at 30Β°C) in the presence of organic solvent, measured by HPLC Tetrahydrofuran (THF) 10% 99.6 98.2 65.3 % 50 mM Tris-HCl 7.5 30Β°C 4 mM Ketoisophorone Levodione 4 mM NADH 180 rpm Classical aqueous control (in %)
DEL267-270 Activity + Stability - Product Formation Product yield after incubaton (4 hours at 30Β°C) in the presence of organic solvent, measured by HPLC Tetrahydrofuran (THF) 20% 99.6 17.3 41.3 % 50 mM Tris-HCl 7.5 30Β°C 4 mM Ketoisophorone Levodione 4 mM NADH 180 rpm Classical aqueous control (in %)
DEL267-270 Activity + Stability - Product Formation Product yield after incubaton (4 hours at 30Β°C) in the presence of organic solvent, measured by HPLC Tetrahydrofuran (THF) 30% 99.6 1.8 12.0 % 50 mM Tris-HCl 7.5 30Β°C 4 mM Ketoisophorone Levodione 4 mM NADH 180 rpm Classical aqueous control (in %)
DEL230-236 Activity + Stability - Product Formation Product yield after incubaton (4 hours at 30Β°C) in the presence of organic solvent, measured by HPLC Tetrahydrofuran (THF) 30% 97.3 1.8 1.3 % 50 mM Tris-HCl 7.5 30Β°C 4 mM Ketoisophorone Levodione 4 mM NADH 180 rpm Classical aqueous control (in %)
DEL230-236 Activity + Stability - Product Formation Product yield after incubaton (4 hours at 30Β°C) in the presence of organic solvent, measured by HPLC Tetrahydrofuran (THF) 20% 97.3 17.3 1.3 % 50 mM Tris-HCl 7.5 30Β°C 4 mM Ketoisophorone Levodione 4 mM NADH 180 rpm Classical aqueous control (in %)
DEL230-236 Activity + Stability - Product Formation Product yield after incubaton (4 hours at 30Β°C) in the presence of organic solvent, measured by HPLC Tetrahydrofuran (THF) 10% 97.3 98.2 11.1 % 50 mM Tris-HCl 7.5 30Β°C 4 mM Ketoisophorone Levodione 4 mM NADH 180 rpm Classical aqueous control (in %)
DEL230-236 Activity + Stability - Product Formation Product yield after incubaton (4 hours at 30Β°C) in the presence of organic solvent, measured by HPLC Tetrahydrofuran (THF) 5% 97.3 90.2 54.7 % 50 mM Tris-HCl 7.5 30Β°C 4 mM Ketoisophorone Levodione 4 mM NADH 180 rpm Classical aqueous control (in %)
DEL230-236 Activity + Stability - Product Formation Product yield after incubaton (4 hours at 30Β°C) in the presence of organic solvent, measured by HPLC Ethyl Acetate 30% 97.3 23.8 16.1 % 50 mM Tris-HCl 7.5 30Β°C 4 mM Ketoisophorone Levodione 4 mM NADH 180 rpm Classical aqueous control (in %)
DEL230-236 Activity + Stability - Product Formation Product yield after incubaton (4 hours at 30Β°C) in the presence of organic solvent, measured by HPLC Ethyl Acetate 20% 97.3 36.3 4.9 % 50 mM Tris-HCl 7.5 30Β°C 4 mM Ketoisophorone Levodione 4 mM NADH 180 rpm Classical aqueous control (in %)
DEL230-236 Activity + Stability - Product Formation Product yield after incubaton (4 hours at 30Β°C) in the presence of organic solvent, measured by HPLC Ethyl Acetate 10% 97.3 65.0 12.1 % 50 mM Tris-HCl 7.5 30Β°C 4 mM Ketoisophorone Levodione 4 mM NADH 180 rpm Classical aqueous control (in %)
DEL230-236 Activity + Stability - Product Formation Product yield after incubaton (4 hours at 30Β°C) in the presence of organic solvent, measured by HPLC Ethyl Acetate 5% 97.3 79.8 35.9 % 50 mM Tris-HCl 7.5 30Β°C 4 mM Ketoisophorone Levodione 4 mM NADH 180 rpm Classical aqueous control (in %)
1 2 3

Mutations in this dataset (3) β€” Sabrina Reich et al. (2014)

WT DEL230-236 DEL267-270

Visualization : Activity + Stability β€” Product Formation

Sabrina Reich et al. (2014)

One bar per measurement. Colour = solvent, shade = solvent volume. β€” β€” β€” Reference value. Hover for details.

Mutation Effect

Mutation impact on enzyme stability and function in the presence of organic solvent: comparison of wild-type and mutant values in identical conditions.

Frieda A. Sorgenfrei et al. (2024) β€” Solvent concentration at 50% protein unfolding may reform enzyme stability ranking and process window identification
Nature Communications  Β· doi:10.1038/s41467-024-49774-0 β†—  Β· Activity - Classical Stability - Tm
150 measurements

Structure

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