Yeast-expressed hydroxynitrile lyase (gene PeHNL) from Passiflora edulis (passion fruit)
Enzyme Description
Sequence
MRNPGASFLIKHFLVLSLFLCAGTAHNPPEIVRHIVFNRYKSQLSQKQIDQIIADYGNLQNIAPEMKEWKWGTDLGPAVEDRADGFTHAYESTFHSVADFLNFFYSPPALEFAKEFFPACEKIVVLNYIINETFPYTWALPNKYVVT
Aem Nuylert et al. (2017)
β Effect of Glycosylation on the Biocatalytic Properties of Hydroxynitrile Lyase from the Passion Fruit, Passiflora edulis : A Comparison of Natural and Recombinant Enzymes
ChemBioChem
Β· doi:10.1002/cbic.201600447 β
Β· Stability - Incubation Specificity - Enantioselectivity
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Database ID
UniProt: A0A1L7NZN4 β
Sequence Annotation
Explicit - Provided
Protein Source
Recombinant, host yeast Pichia pastoris GS115
Experimental Data (24 measurements)
24 measurements β page 1 of 2
| Mutation | Property | Assay | Solvent | Solvent Volume | Aqueous Reference | WT Reference | Measured Value | Units | Solution | pH | Temperature | Substrate(s) | Product(s) | Cofactor(s) | Shaking | Comments |
|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
| WT | Specificity - Enantioselectivity | Enantiomeric excess (toward R form) after 12 hours incubation at 10Β°C in the presence of organic solvent, measured by HPLC | n-Hexane | 50% | 50.4 | β | 32.2 | % | 400 mM citrate buffer | 4 | 10Β°C | 900 mM Acetone cyanohydrin, 250 mM Benzaldehyde | (R)-Mandelonitrile | β | β | Classical aqueous control (in %), with activity measured in the presence of organic solvent before and after incubation for determining 100% value | Acetone cyanohydrin forms HCN |
| WT | Specificity - Enantioselectivity | Enantiomeric excess (toward R form) after 12 hours incubation at 10Β°C in the presence of organic solvent, measured by HPLC | Butyl Ether | 50% | 50.4 | β | 94.7 | % | 400 mM citrate buffer | 4 | 10Β°C | 900 mM Acetone cyanohydrin, 250 mM Benzaldehyde | (R)-Mandelonitrile | β | β | Classical aqueous control (in %), with activity measured in the presence of organic solvent before and after incubation for determining 100% value | Acetone cyanohydrin forms HCN |
| WT | Specificity - Enantioselectivity | Enantiomeric excess (toward R form) after 12 hours incubation at 10Β°C in the presence of organic solvent, measured by HPLC | Diisopropyl Ether | 50% | 50.4 | β | 98.1 | % | 400 mM citrate buffer | 4 | 10Β°C | 900 mM Acetone cyanohydrin, 250 mM Benzaldehyde | (R)-Mandelonitrile | β | β | Classical aqueous control (in %), with activity measured in the presence of organic solvent before and after incubation for determining 100% value | Acetone cyanohydrin forms HCN |
| WT | Specificity - Enantioselectivity | Enantiomeric excess (toward R form) after 12 hours incubation at 10Β°C in the presence of organic solvent, measured by HPLC | Ethyl Acetate | 50% | 50.4 | β | 95.9 | % | 400 mM citrate buffer | 4 | 10Β°C | 900 mM Acetone cyanohydrin, 250 mM Benzaldehyde | (R)-Mandelonitrile | β | β | Classical aqueous control (in %), with activity measured in the presence of organic solvent before and after incubation for determining 100% value | Acetone cyanohydrin forms HCN |
| WT | Specificity - Enantioselectivity | Enantiomeric excess (toward R form) after 12 hours incubation at 10Β°C in the presence of organic solvent, measured by HPLC | Diethyl Ether | 50% | 50.4 | β | 95.4 | % | 400 mM citrate buffer | 4 | 10Β°C | 900 mM Acetone cyanohydrin, 250 mM Benzaldehyde | (R)-Mandelonitrile | β | β | Classical aqueous control (in %), with activity measured in the presence of organic solvent before and after incubation for determining 100% value | Acetone cyanohydrin forms HCN |
| WT | Specificity - Enantioselectivity | Enantiomeric excess (toward R form) after 12 hours incubation at 10Β°C in the presence of organic solvent, measured by HPLC | tert-Butyl Methyl Ether (MTBE) | 50% | 50.4 | β | 94.4 | % | 400 mM citrate buffer | 4 | 10Β°C | 900 mM Acetone cyanohydrin, 250 mM Benzaldehyde | (R)-Mandelonitrile | β | β | Classical aqueous control (in %), with activity measured in the presence of organic solvent before and after incubation for determining 100% value | Acetone cyanohydrin forms HCN |
| WT | Stability - Incubation | Activity after incubation (12 hours at 10Β°C), measured by HPLC in presence of organic solvent | Ethyl Acetate | 50% | 90.4 | β | 55.9 | % | Incubation: 400 mM citrate buffer, Assay: 400 mM citrate buffer | Incubation: 4, Assay: 4 | ? | 900 mM Acetone cyanohydrin, 250 mM Benzaldehyde | (R)-Mandelonitrile | β | β | Water-incubated control (in %), with activity measured in the presence of organic solvent before and after incubation for determining 100% value | Acetone cyanohydrin forms HCN |
| WT | Stability - Incubation | Activity after incubation (12 hours at 10Β°C), measured by HPLC in presence of organic solvent | Diethyl Ether | 50% | 90.4 | β | 82.6 | % | Incubation: 400 mM citrate buffer, Assay: 400 mM citrate buffer | Incubation: 4, Assay: 4 | ? | 900 mM Acetone cyanohydrin, 250 mM Benzaldehyde | (R)-Mandelonitrile | β | β | Water-incubated control (in %), with activity measured in the presence of organic solvent before and after incubation for determining 100% value | Acetone cyanohydrin forms HCN |
| WT | Stability - Incubation | Activity after incubation (12 hours at 10Β°C), measured by HPLC in presence of organic solvent | tert-Butyl Methyl Ether (MTBE) | 50% | 90.4 | β | 65.3 | % | Incubation: 400 mM citrate buffer, Assay: 400 mM citrate buffer | Incubation: 4, Assay: 4 | ? | 900 mM Acetone cyanohydrin, 250 mM Benzaldehyde | (R)-Mandelonitrile | β | β | Water-incubated control (in %), with activity measured in the presence of organic solvent before and after incubation for determining 100% value | Acetone cyanohydrin forms HCN |
| WT | Stability - Incubation | Activity after incubation (12 hours at 10Β°C), measured by HPLC in presence of organic solvent | Diisopropyl Ether | 50% | 90.4 | β | 93.4 | % | Incubation: 400 mM citrate buffer, Assay: 400 mM citrate buffer | Incubation: 4, Assay: 4 | ? | 900 mM Acetone cyanohydrin, 250 mM Benzaldehyde | (R)-Mandelonitrile | β | β | Water-incubated control (in %), with activity measured in the presence of organic solvent before and after incubation for determining 100% value | Acetone cyanohydrin forms HCN |
| WT | Stability - Incubation | Activity after incubation (12 hours at 10Β°C), measured by HPLC in presence of organic solvent | Butyl Ether | 50% | 90.4 | β | 65.0 | % | Incubation: 400 mM citrate buffer, Assay: 400 mM citrate buffer | Incubation: 4, Assay: 4 | ? | 900 mM Acetone cyanohydrin, 250 mM Benzaldehyde | (R)-Mandelonitrile | β | β | Water-incubated control (in %), with activity measured in the presence of organic solvent before and after incubation for determining 100% value | Acetone cyanohydrin forms HCN |
| WT | Stability - Incubation | Activity after incubation (12 hours at 10Β°C), measured by HPLC in presence of organic solvent | n-Hexane | 50% | 90.4 | β | 72.4 | % | Incubation: 400 mM citrate buffer, Assay: 400 mM citrate buffer | Incubation: 4, Assay: 4 | ? | 900 mM Acetone cyanohydrin, 250 mM Benzaldehyde | (R)-Mandelonitrile | β | β | Water-incubated control (in %), with activity measured in the presence of organic solvent before and after incubation for determining 100% value | Acetone cyanohydrin forms HCN |
| N105Q | Specificity - Enantioselectivity | Enantiomeric excess (toward R form) after 12 hours incubation at 10Β°C in the presence of organic solvent, measured by HPLC | n-Hexane | 50% | 53.3 | 32.2 | 27.8 | % | 400 mM citrate buffer | 4 | 10Β°C | 900 mM Acetone cyanohydrin, 250 mM Benzaldehyde | (R)-Mandelonitrile | β | β | Classical aqueous control (in %), with activity measured in the presence of organic solvent before and after incubation for determining 100% value | Acetone cyanohydrin forms HCN |
| N105Q | Specificity - Enantioselectivity | Enantiomeric excess (toward R form) after 12 hours incubation at 10Β°C in the presence of organic solvent, measured by HPLC | Butyl Ether | 50% | 53.3 | 94.7 | 94.9 | % | 400 mM citrate buffer | 4 | 10Β°C | 900 mM Acetone cyanohydrin, 250 mM Benzaldehyde | (R)-Mandelonitrile | β | β | Classical aqueous control (in %), with activity measured in the presence of organic solvent before and after incubation for determining 100% value | Acetone cyanohydrin forms HCN |
| N105Q | Specificity - Enantioselectivity | Enantiomeric excess (toward R form) after 12 hours incubation at 10Β°C in the presence of organic solvent, measured by HPLC | Diisopropyl Ether | 50% | 53.3 | 98.1 | 97.6 | % | 400 mM citrate buffer | 4 | 10Β°C | 900 mM Acetone cyanohydrin, 250 mM Benzaldehyde | (R)-Mandelonitrile | β | β | Classical aqueous control (in %), with activity measured in the presence of organic solvent before and after incubation for determining 100% value | Acetone cyanohydrin forms HCN |
| N105Q | Specificity - Enantioselectivity | Enantiomeric excess (toward R form) after 12 hours incubation at 10Β°C in the presence of organic solvent, measured by HPLC | tert-Butyl Methyl Ether (MTBE) | 50% | 53.3 | 94.4 | 94.7 | % | 400 mM citrate buffer | 4 | 10Β°C | 900 mM Acetone cyanohydrin, 250 mM Benzaldehyde | (R)-Mandelonitrile | β | β | Classical aqueous control (in %), with activity measured in the presence of organic solvent before and after incubation for determining 100% value | Acetone cyanohydrin forms HCN |
| N105Q | Specificity - Enantioselectivity | Enantiomeric excess (toward R form) after 12 hours incubation at 10Β°C in the presence of organic solvent, measured by HPLC | Diethyl Ether | 50% | 53.3 | 95.4 | 94.7 | % | 400 mM citrate buffer | 4 | 10Β°C | 900 mM Acetone cyanohydrin, 250 mM Benzaldehyde | (R)-Mandelonitrile | β | β | Classical aqueous control (in %), with activity measured in the presence of organic solvent before and after incubation for determining 100% value | Acetone cyanohydrin forms HCN |
| N105Q | Specificity - Enantioselectivity | Enantiomeric excess (toward R form) after 12 hours incubation at 10Β°C in the presence of organic solvent, measured by HPLC | Ethyl Acetate | 50% | 53.3 | 95.9 | 94.2 | % | 400 mM citrate buffer | 4 | 10Β°C | 900 mM Acetone cyanohydrin, 250 mM Benzaldehyde | (R)-Mandelonitrile | β | β | Classical aqueous control (in %), with activity measured in the presence of organic solvent before and after incubation for determining 100% value | Acetone cyanohydrin forms HCN |
| N105Q | Stability - Incubation | Activity after incubation (12 hours at 10Β°C), measured by HPLC in presence of organic solvent | n-Hexane | 50% | 55.4 | 72.4 | 67.8 | % | Incubation: 400 mM citrate buffer, Assay: 400 mM citrate buffer | Incubation: 4, Assay: 4 | ? | 900 mM Acetone cyanohydrin, 250 mM Benzaldehyde | (R)-Mandelonitrile | β | β | Water-incubated control (in %), with activity measured in the presence of organic solvent before and after incubation for determining 100% value | Acetone cyanohydrin forms HCN |
| N105Q | Stability - Incubation | Activity after incubation (12 hours at 10Β°C), measured by HPLC in presence of organic solvent | Butyl Ether | 50% | 55.4 | 65.0 | 62.5 | % | Incubation: 400 mM citrate buffer, Assay: 400 mM citrate buffer | Incubation: 4, Assay: 4 | ? | 900 mM Acetone cyanohydrin, 250 mM Benzaldehyde | (R)-Mandelonitrile | β | β | Water-incubated control (in %), with activity measured in the presence of organic solvent before and after incubation for determining 100% value | Acetone cyanohydrin forms HCN |
Mutations in this dataset (2)
WT
N105Q
Visualization : Stability β Incubation
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.
Visualization : Specificity β Enantioselectivity
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.
Structure
π§¬
No structure available for this protein.