E.Coli-expressed hydroxynitrile lyase (gene PeHNL) from Passiflora edulis (passion fruit)

Enzyme Description

Species

Extremophile

EC Number

4.1.2.10 ↗

Sequence

Length: 147 amino acids

MRNPGASFLIKHFLVLSLFLCAGTAHNPPEIVRHIVFNRYKSQLSQKQIDQIIADYGNLQNIAPEMKEWKWGTDLGPAVEDRADGFTHAYESTFHSVADFLNFFYSPPALEFAKEFFPACEKIVVLNYIINETFPYTWALPNKYVVT

Aem Nuylert et al. (2017) — Effect of Glycosylation on the Biocatalytic Properties of Hydroxynitrile Lyase from the Passion Fruit, Passiflora edulis : A Comparison of Natural and Recombinant Enzymes

ChemBioChem · doi:10.1002/cbic.201600447 ↗ · Stability - Incubation Specificity - Enantioselectivity

12 measurements ▼

Database ID

UniProt: A0A1L7NZN4 ↗

Sequence Annotation

Explicit - Provided

Protein Source

Recombinant, host bacterium Escherichia coli BL21

Experimental Data (12 measurements)

12 measurements

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 R form) after 12 hours incubation at 10°C in the presence of organic solvent, measured by HPLC	Ethyl Acetate	50%	51.8	84.5	%	400 mM citrate buffer	4	10°C	900 mM Acetone cyanohydrin, 250 mM Benzaldehyde	(R)-Mandelonitrile	—	—	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
Specificity - Enantioselectivity	Enantiomeric excess (toward R form) after 12 hours incubation at 10°C in the presence of organic solvent, measured by HPLC	Diethyl Ether	50%	51.8	96.6	%	400 mM citrate buffer	4	10°C	900 mM Acetone cyanohydrin, 250 mM Benzaldehyde	(R)-Mandelonitrile	—	—	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
Specificity - Enantioselectivity	Enantiomeric excess (toward R form) after 12 hours incubation at 10°C in the presence of organic solvent, measured by HPLC	tert-Butyl Methyl Ether (MTBE)	50%	51.8	94.4	%	400 mM citrate buffer	4	10°C	900 mM Acetone cyanohydrin, 250 mM Benzaldehyde	(R)-Mandelonitrile	—	—	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
Specificity - Enantioselectivity	Enantiomeric excess (toward R form) after 12 hours incubation at 10°C in the presence of organic solvent, measured by HPLC	Diisopropyl Ether	50%	51.8	94.9	%	400 mM citrate buffer	4	10°C	900 mM Acetone cyanohydrin, 250 mM Benzaldehyde	(R)-Mandelonitrile	—	—	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
Specificity - Enantioselectivity	Enantiomeric excess (toward R form) after 12 hours incubation at 10°C in the presence of organic solvent, measured by HPLC	Butyl Ether	50%	51.8	91.3	%	400 mM citrate buffer	4	10°C	900 mM Acetone cyanohydrin, 250 mM Benzaldehyde	(R)-Mandelonitrile	—	—	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
Specificity - Enantioselectivity	Enantiomeric excess (toward R form) after 12 hours incubation at 10°C in the presence of organic solvent, measured by HPLC	n-Hexane	50%	51.8	31.2	%	400 mM citrate buffer	4	10°C	900 mM Acetone cyanohydrin, 250 mM Benzaldehyde	(R)-Mandelonitrile	—	—	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
Stability - Incubation	Activity after incubation (12 hours at 10°C), measured by HPLC in presence of organic solvent	Ethyl Acetate	50%	53.4	1.9	%	Incubation: 400 mM citrate buffer, Assay: 400 mM citrate buffer	Incubation: 4, Assay: 4	?	900 mM Acetone cyanohydrin, 250 mM Benzaldehyde	(R)-Mandelonitrile	—	—	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
Stability - Incubation	Activity after incubation (12 hours at 10°C), measured by HPLC in presence of organic solvent	Diethyl Ether	50%	53.4	4.1	%	Incubation: 400 mM citrate buffer, Assay: 400 mM citrate buffer	Incubation: 4, Assay: 4	?	900 mM Acetone cyanohydrin, 250 mM Benzaldehyde	(R)-Mandelonitrile	—	—	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
Stability - Incubation	Activity after incubation (12 hours at 10°C), measured by HPLC in presence of organic solvent	tert-Butyl Methyl Ether (MTBE)	50%	53.4	25.3	%	Incubation: 400 mM citrate buffer, Assay: 400 mM citrate buffer	Incubation: 4, Assay: 4	?	900 mM Acetone cyanohydrin, 250 mM Benzaldehyde	(R)-Mandelonitrile	—	—	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
Stability - Incubation	Activity after incubation (12 hours at 10°C), measured by HPLC in presence of organic solvent	Diisopropyl Ether	50%	53.4	20.1	%	Incubation: 400 mM citrate buffer, Assay: 400 mM citrate buffer	Incubation: 4, Assay: 4	?	900 mM Acetone cyanohydrin, 250 mM Benzaldehyde	(R)-Mandelonitrile	—	—	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
Stability - Incubation	Activity after incubation (12 hours at 10°C), measured by HPLC in presence of organic solvent	Butyl Ether	50%	53.4	52.9	%	Incubation: 400 mM citrate buffer, Assay: 400 mM citrate buffer	Incubation: 4, Assay: 4	?	900 mM Acetone cyanohydrin, 250 mM Benzaldehyde	(R)-Mandelonitrile	—	—	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
Stability - Incubation	Activity after incubation (12 hours at 10°C), measured by HPLC in presence of organic solvent	n-Hexane	50%	53.4	69.4	%	Incubation: 400 mM citrate buffer, Assay: 400 mM citrate buffer	Incubation: 4, Assay: 4	?	900 mM Acetone cyanohydrin, 250 mM Benzaldehyde	(R)-Mandelonitrile	—	—	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	Specificity - Enantioselectivity	Enantiomeric excess (toward R form) after 12 hours incubation at 10°C in the presence of organic solvent, measured by HPLC	Ethyl Acetate	50%	51.8	84.5	%	400 mM citrate buffer	4	10°C	900 mM Acetone cyanohydrin, 250 mM Benzaldehyde	(R)-Mandelonitrile	—	—	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 10°C in the presence of organic solvent, measured by HPLC	Diethyl Ether	50%	51.8	96.6	%	400 mM citrate buffer	4	10°C	900 mM Acetone cyanohydrin, 250 mM Benzaldehyde	(R)-Mandelonitrile	—	—	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 10°C in the presence of organic solvent, measured by HPLC	tert-Butyl Methyl Ether (MTBE)	50%	51.8	94.4	%	400 mM citrate buffer	4	10°C	900 mM Acetone cyanohydrin, 250 mM Benzaldehyde	(R)-Mandelonitrile	—	—	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 10°C in the presence of organic solvent, measured by HPLC	Diisopropyl Ether	50%	51.8	94.9	%	400 mM citrate buffer	4	10°C	900 mM Acetone cyanohydrin, 250 mM Benzaldehyde	(R)-Mandelonitrile	—	—	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 10°C in the presence of organic solvent, measured by HPLC	Butyl Ether	50%	51.8	91.3	%	400 mM citrate buffer	4	10°C	900 mM Acetone cyanohydrin, 250 mM Benzaldehyde	(R)-Mandelonitrile	—	—	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 10°C in the presence of organic solvent, measured by HPLC	n-Hexane	50%	51.8	31.2	%	400 mM citrate buffer	4	10°C	900 mM Acetone cyanohydrin, 250 mM Benzaldehyde	(R)-Mandelonitrile	—	—	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 10°C), measured by HPLC in presence of organic solvent	Ethyl Acetate	50%	53.4	1.9	%	Incubation: 400 mM citrate buffer, Assay: 400 mM citrate buffer	Incubation: 4, Assay: 4	?	900 mM Acetone cyanohydrin, 250 mM Benzaldehyde	(R)-Mandelonitrile	—	—	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 10°C), measured by HPLC in presence of organic solvent	Diethyl Ether	50%	53.4	4.1	%	Incubation: 400 mM citrate buffer, Assay: 400 mM citrate buffer	Incubation: 4, Assay: 4	?	900 mM Acetone cyanohydrin, 250 mM Benzaldehyde	(R)-Mandelonitrile	—	—	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 10°C), measured by HPLC in presence of organic solvent	tert-Butyl Methyl Ether (MTBE)	50%	53.4	25.3	%	Incubation: 400 mM citrate buffer, Assay: 400 mM citrate buffer	Incubation: 4, Assay: 4	?	900 mM Acetone cyanohydrin, 250 mM Benzaldehyde	(R)-Mandelonitrile	—	—	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 10°C), measured by HPLC in presence of organic solvent	Diisopropyl Ether	50%	53.4	20.1	%	Incubation: 400 mM citrate buffer, Assay: 400 mM citrate buffer	Incubation: 4, Assay: 4	?	900 mM Acetone cyanohydrin, 250 mM Benzaldehyde	(R)-Mandelonitrile	—	—	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 10°C), measured by HPLC in presence of organic solvent	Butyl Ether	50%	53.4	52.9	%	Incubation: 400 mM citrate buffer, Assay: 400 mM citrate buffer	Incubation: 4, Assay: 4	?	900 mM Acetone cyanohydrin, 250 mM Benzaldehyde	(R)-Mandelonitrile	—	—	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 10°C), measured by HPLC in presence of organic solvent	n-Hexane	50%	53.4	69.4	%	Incubation: 400 mM citrate buffer, Assay: 400 mM citrate buffer	Incubation: 4, Assay: 4	?	900 mM Acetone cyanohydrin, 250 mM Benzaldehyde	(R)-Mandelonitrile	—	—	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

Visualization : Stability — Incubation

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

Visualization : Specificity — Enantioselectivity