Yeast-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

24 measurements ▼

Database ID

UniProt: A0A1L7NZN4 ↗

Sequence Annotation

Explicit - Provided

Protein Source

Recombinant, host yeast Pichia pastoris GS115

Experimental Data (24 measurements)

24 measurements — page 2 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
N105Q	Stability - Incubation	Activity after incubation (12 hours at 10°C), measured by HPLC in presence of organic solvent	Diisopropyl Ether	50%	55.4	93.4	86.2	%	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
N105Q	Stability - Incubation	Activity after incubation (12 hours at 10°C), measured by HPLC in presence of organic solvent	tert-Butyl Methyl Ether (MTBE)	50%	55.4	65.3	31.1	%	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
N105Q	Stability - Incubation	Activity after incubation (12 hours at 10°C), measured by HPLC in presence of organic solvent	Diethyl Ether	50%	55.4	82.6	75.2	%	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
N105Q	Stability - Incubation	Activity after incubation (12 hours at 10°C), measured by HPLC in presence of organic solvent	Ethyl Acetate	50%	55.4	55.9	0.0	%	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	n-Hexane	50%	50.4	—	32.2	%	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	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%	50.4	—	94.7	%	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%	50.4	—	98.1	%	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	Ethyl Acetate	50%	50.4	—	95.9	%	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	Diethyl Ether	50%	50.4	—	95.4	%	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	tert-Butyl Methyl Ether (MTBE)	50%	50.4	—	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	Stability - Incubation	Activity after incubation (12 hours at 10°C), measured by HPLC in presence of organic solvent	Ethyl Acetate	50%	90.4	—	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	—	—	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%	90.4	—	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	—	—	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%	90.4	—	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	—	—	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%	90.4	—	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	—	—	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%	90.4	—	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	—	—	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%	90.4	—	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	—	—	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 10°C in the presence of organic solvent, measured by HPLC	n-Hexane	50%	53.3	32.2	27.8	%	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
N105Q	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%	53.3	94.7	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
N105Q	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%	53.3	98.1	97.6	%	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
N105Q	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%	53.3	94.4	94.7	%	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
N105Q	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%	53.3	95.4	94.7	%	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
N105Q	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%	53.3	95.9	94.2	%	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
N105Q	Stability - Incubation	Activity after incubation (12 hours at 10°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	—	—	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 10°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	—	—	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 10°C), measured by HPLC in presence of organic solvent	Diisopropyl Ether	50%	55.4	93.4	86.2	%	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
N105Q	Stability - Incubation	Activity after incubation (12 hours at 10°C), measured by HPLC in presence of organic solvent	tert-Butyl Methyl Ether (MTBE)	50%	55.4	65.3	31.1	%	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
N105Q	Stability - Incubation	Activity after incubation (12 hours at 10°C), measured by HPLC in presence of organic solvent	Diethyl Ether	50%	55.4	82.6	75.2	%	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
N105Q	Stability - Incubation	Activity after incubation (12 hours at 10°C), measured by HPLC in presence of organic solvent	Ethyl Acetate	50%	55.4	55.9	0.0	%	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

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Mutations in this dataset (2)

WT N105Q

Visualization : Stability — Incubation

One bar per measurement. Colour = solvent, shade = solvent volume. — — — 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 — Enantioselectivity

One bar per measurement. Colour = solvent, shade = solvent volume. — — — 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

🧬

No structure available for this protein.

Yeast-expressed hydroxynitrile lyase (gene PeHNL) from Passiflora edulis (passion fruit)

Enzyme Description

Sequence

Experimental Data (24 measurements)

Visualization : Stability — Incubation

Mutation Effect

Visualization : Specificity — Enantioselectivity

Mutation Effect

Structure

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