Pectate lyase from Aspergillus luchuensis var. saitoi

Enzyme Description

Species
Aspergillus luchuensis var. saitoi β†—
Extremophile
No
EC Number
4.2.2.2 β†—

Sequence

Length: 323 amino acids
MTNFKWIVAAAGLLSGQVLAAPTATSTHAKRATVSDAAFGYASMNGGTTGGAGGTTTTVSSYAAFTSAVSGDDAKVVYVDGTIKETADQVKIGSNTSIIGKDANAILEGFGVLVKEKENVIIRNLGVTKVLADNGDAIGIQYSNNVWIDHCDVSSDRDHDKDYYDGLIDITHGSDYVTVSNTFIHDHWKASLVGHSVSNEDEDSGHLTVTYANNYWYNVNSRAPSFRFGTGHIYNSYYLDVSDGINTRDGAQLLVESNQFVDSKKALYSTDDGYAVSNDNDFGDAKSTAEEGTLTSMPYDYTLLGSANVKAAVVGTAGQTLTF
Junya Kamijo et al. (2019) β€” Identification and characterization of a thermostable pectate lyase from Aspergillus luchuensis var. saitoi
Food Chemistry  Β· doi:10.1016/j.foodchem.2018.10.059 β†—  Β· Stability - Incubation
64 measurements ▼
Database ID
β€”
Sequence Annotation
Explicit - Provided
Protein Source
Recombinant, host yeast Pichia pastoris KM71H

Experimental Data (64 measurements)

64 measurements β€” page 2 of 4
Mutation Property Assay Solvent Solvent Volume Aqueous ReferenceWT Reference Measured Value Units Solution pH Temperature Substrate(s) Product(s) Cofactor(s) Shaking Comments
I140V/V197D Stability - Incubation Activity after incubation (1 hour at 30Β°C), measured by absorbance spectrophotometry (unsaturated bonds absorbance measurement, 235 nm) Isopropanol 10% 100 85 79 % Incubation: 50 mM Tris-HCl buffer, Assay: 50 mM Tris-HCl buffer, 0.3 MM CaCl2 Incubation: 8, Assay: 8 Incubation: 30Β°C, Assay: 30Β°C 0.5% Polygalacturonic acid Ξ”4,5-unsaturated oligogalacturonic acids 0.3 mM Ca²⁺ β€” Water-incubated control (in %), assay in aqueous phase | Uncertainty about whether assay really is in aqueous phase/uncertainty about EC number and reaction buffer
V197D Stability - Incubation Activity after incubation (1 hour at 30Β°C), measured by absorbance spectrophotometry (unsaturated bonds absorbance measurement, 235 nm) Methanol 40% 100 22 19 % Incubation: 50 mM Tris-HCl buffer, Assay: 50 mM Tris-HCl buffer, 0.3 MM CaCl2 Incubation: 8, Assay: 8 Incubation: 30Β°C, Assay: 30Β°C 0.5% Polygalacturonic acid Ξ”4,5-unsaturated oligogalacturonic acids 0.3 mM Ca²⁺ β€” Water-incubated control (in %), assay in aqueous phase | Uncertainty about whether assay really is in aqueous phase/uncertainty about EC number and reaction buffer
I140V Stability - Incubation Activity after incubation (1 hour at 30Β°C), measured by absorbance spectrophotometry (unsaturated bonds absorbance measurement, 235 nm) Methanol 40% 100 22 24 % Incubation: 50 mM Tris-HCl buffer, Assay: 50 mM Tris-HCl buffer, 0.3 MM CaCl2 Incubation: 8, Assay: 8 Incubation: 30Β°C, Assay: 30Β°C 0.5% Polygalacturonic acid Ξ”4,5-unsaturated oligogalacturonic acids 0.3 mM Ca²⁺ β€” Water-incubated control (in %), assay in aqueous phase | Uncertainty about whether assay really is in aqueous phase/uncertainty about EC number and reaction buffer
I140V Stability - Incubation Activity after incubation (1 hour at 30Β°C), measured by absorbance spectrophotometry (unsaturated bonds absorbance measurement, 235 nm) Isopropanol 20% 100 25 25 % Incubation: 50 mM Tris-HCl buffer, Assay: 50 mM Tris-HCl buffer, 0.3 MM CaCl2 Incubation: 8, Assay: 8 Incubation: 30Β°C, Assay: 30Β°C 0.5% Polygalacturonic acid Ξ”4,5-unsaturated oligogalacturonic acids 0.3 mM Ca²⁺ β€” Water-incubated control (in %), assay in aqueous phase | Uncertainty about whether assay really is in aqueous phase/uncertainty about EC number and reaction buffer
V197D Stability - Incubation Activity after incubation (1 hour at 30Β°C), measured by absorbance spectrophotometry (unsaturated bonds absorbance measurement, 235 nm) Isopropanol 20% 100 25 29 % Incubation: 50 mM Tris-HCl buffer, Assay: 50 mM Tris-HCl buffer, 0.3 MM CaCl2 Incubation: 8, Assay: 8 Incubation: 30Β°C, Assay: 30Β°C 0.5% Polygalacturonic acid Ξ”4,5-unsaturated oligogalacturonic acids 0.3 mM Ca²⁺ β€” Water-incubated control (in %), assay in aqueous phase | Uncertainty about whether assay really is in aqueous phase/uncertainty about EC number and reaction buffer
I140V/V197D Stability - Incubation Activity after incubation (1 hour at 30Β°C), measured by absorbance spectrophotometry (unsaturated bonds absorbance measurement, 235 nm) Isopropanol 20% 100 25 30 % Incubation: 50 mM Tris-HCl buffer, Assay: 50 mM Tris-HCl buffer, 0.3 MM CaCl2 Incubation: 8, Assay: 8 Incubation: 30Β°C, Assay: 30Β°C 0.5% Polygalacturonic acid Ξ”4,5-unsaturated oligogalacturonic acids 0.3 mM Ca²⁺ β€” Water-incubated control (in %), assay in aqueous phase | Uncertainty about whether assay really is in aqueous phase/uncertainty about EC number and reaction buffer
I140V Stability - Incubation Activity after incubation (1 hour at 30Β°C), measured by absorbance spectrophotometry (unsaturated bonds absorbance measurement, 235 nm) Isopropanol 40% 100 19 17 % Incubation: 50 mM Tris-HCl buffer, Assay: 50 mM Tris-HCl buffer, 0.3 MM CaCl2 Incubation: 8, Assay: 8 Incubation: 30Β°C, Assay: 30Β°C 0.5% Polygalacturonic acid Ξ”4,5-unsaturated oligogalacturonic acids 0.3 mM Ca²⁺ β€” Water-incubated control (in %), assay in aqueous phase | Uncertainty about whether assay really is in aqueous phase/uncertainty about EC number and reaction buffer
I140V/V197D Stability - Incubation Activity after incubation (1 hour at 30Β°C), measured by absorbance spectrophotometry (unsaturated bonds absorbance measurement, 235 nm) Methanol 10% 100 92 88 % Incubation: 50 mM Tris-HCl buffer, Assay: 50 mM Tris-HCl buffer, 0.3 MM CaCl2 Incubation: 8, Assay: 8 Incubation: 30Β°C, Assay: 30Β°C 0.5% Polygalacturonic acid Ξ”4,5-unsaturated oligogalacturonic acids 0.3 mM Ca²⁺ β€” Water-incubated control (in %), assay in aqueous phase | Uncertainty about whether assay really is in aqueous phase/uncertainty about EC number and reaction buffer
V197D Stability - Incubation Activity after incubation (1 hour at 30Β°C), measured by absorbance spectrophotometry (unsaturated bonds absorbance measurement, 235 nm) Isopropanol 40% 100 19 19 % Incubation: 50 mM Tris-HCl buffer, Assay: 50 mM Tris-HCl buffer, 0.3 MM CaCl2 Incubation: 8, Assay: 8 Incubation: 30Β°C, Assay: 30Β°C 0.5% Polygalacturonic acid Ξ”4,5-unsaturated oligogalacturonic acids 0.3 mM Ca²⁺ β€” Water-incubated control (in %), assay in aqueous phase | Uncertainty about whether assay really is in aqueous phase/uncertainty about EC number and reaction buffer
I140V/V197D Stability - Incubation Activity after incubation (1 hour at 30Β°C), measured by absorbance spectrophotometry (unsaturated bonds absorbance measurement, 235 nm) Isopropanol 40% 100 19 18 % Incubation: 50 mM Tris-HCl buffer, Assay: 50 mM Tris-HCl buffer, 0.3 MM CaCl2 Incubation: 8, Assay: 8 Incubation: 30Β°C, Assay: 30Β°C 0.5% Polygalacturonic acid Ξ”4,5-unsaturated oligogalacturonic acids 0.3 mM Ca²⁺ β€” Water-incubated control (in %), assay in aqueous phase | Uncertainty about whether assay really is in aqueous phase/uncertainty about EC number and reaction buffer
V197D Stability - Incubation Activity after incubation (1 hour at 30Β°C), measured by absorbance spectrophotometry (unsaturated bonds absorbance measurement, 235 nm) Methanol 20% 100 62 59 % Incubation: 50 mM Tris-HCl buffer, Assay: 50 mM Tris-HCl buffer, 0.3 MM CaCl2 Incubation: 8, Assay: 8 Incubation: 30Β°C, Assay: 30Β°C 0.5% Polygalacturonic acid Ξ”4,5-unsaturated oligogalacturonic acids 0.3 mM Ca²⁺ β€” Water-incubated control (in %), assay in aqueous phase | Uncertainty about whether assay really is in aqueous phase/uncertainty about EC number and reaction buffer
I140V Stability - Incubation Activity after incubation (1 hour at 30Β°C), measured by absorbance spectrophotometry (unsaturated bonds absorbance measurement, 235 nm) Methanol 5% 100 120 110 % Incubation: 50 mM Tris-HCl buffer, Assay: 50 mM Tris-HCl buffer, 0.3 MM CaCl2 Incubation: 8, Assay: 8 Incubation: 30Β°C, Assay: 30Β°C 0.5% Polygalacturonic acid Ξ”4,5-unsaturated oligogalacturonic acids 0.3 mM Ca²⁺ β€” Water-incubated control (in %), assay in aqueous phase | Uncertainty about whether assay really is in aqueous phase/uncertainty about EC number and reaction buffer
V197D Stability - Incubation Activity after incubation (1 hour at 30Β°C), measured by absorbance spectrophotometry (unsaturated bonds absorbance measurement, 235 nm) Methanol 5% 100 120 116 % Incubation: 50 mM Tris-HCl buffer, Assay: 50 mM Tris-HCl buffer, 0.3 MM CaCl2 Incubation: 8, Assay: 8 Incubation: 30Β°C, Assay: 30Β°C 0.5% Polygalacturonic acid Ξ”4,5-unsaturated oligogalacturonic acids 0.3 mM Ca²⁺ β€” Water-incubated control (in %), assay in aqueous phase | Uncertainty about whether assay really is in aqueous phase/uncertainty about EC number and reaction buffer
I140V/V197D Stability - Incubation Activity after incubation (1 hour at 30Β°C), measured by absorbance spectrophotometry (unsaturated bonds absorbance measurement, 235 nm) Methanol 5% 100 120 120 % Incubation: 50 mM Tris-HCl buffer, Assay: 50 mM Tris-HCl buffer, 0.3 MM CaCl2 Incubation: 8, Assay: 8 Incubation: 30Β°C, Assay: 30Β°C 0.5% Polygalacturonic acid Ξ”4,5-unsaturated oligogalacturonic acids 0.3 mM Ca²⁺ β€” Water-incubated control (in %), assay in aqueous phase | Uncertainty about whether assay really is in aqueous phase/uncertainty about EC number and reaction buffer
I140V Stability - Incubation Activity after incubation (1 hour at 30Β°C), measured by absorbance spectrophotometry (unsaturated bonds absorbance measurement, 235 nm) Methanol 20% 100 62 63 % Incubation: 50 mM Tris-HCl buffer, Assay: 50 mM Tris-HCl buffer, 0.3 MM CaCl2 Incubation: 8, Assay: 8 Incubation: 30Β°C, Assay: 30Β°C 0.5% Polygalacturonic acid Ξ”4,5-unsaturated oligogalacturonic acids 0.3 mM Ca²⁺ β€” Water-incubated control (in %), assay in aqueous phase | Uncertainty about whether assay really is in aqueous phase/uncertainty about EC number and reaction buffer
I140V Stability - Incubation Activity after incubation (1 hour at 30Β°C), measured by absorbance spectrophotometry (unsaturated bonds absorbance measurement, 235 nm) Methanol 10% 100 92 87 % Incubation: 50 mM Tris-HCl buffer, Assay: 50 mM Tris-HCl buffer, 0.3 MM CaCl2 Incubation: 8, Assay: 8 Incubation: 30Β°C, Assay: 30Β°C 0.5% Polygalacturonic acid Ξ”4,5-unsaturated oligogalacturonic acids 0.3 mM Ca²⁺ β€” Water-incubated control (in %), assay in aqueous phase | Uncertainty about whether assay really is in aqueous phase/uncertainty about EC number and reaction buffer
V197D Stability - Incubation Activity after incubation (1 hour at 30Β°C), measured by absorbance spectrophotometry (unsaturated bonds absorbance measurement, 235 nm) Methanol 10% 100 92 88 % Incubation: 50 mM Tris-HCl buffer, Assay: 50 mM Tris-HCl buffer, 0.3 MM CaCl2 Incubation: 8, Assay: 8 Incubation: 30Β°C, Assay: 30Β°C 0.5% Polygalacturonic acid Ξ”4,5-unsaturated oligogalacturonic acids 0.3 mM Ca²⁺ β€” Water-incubated control (in %), assay in aqueous phase | Uncertainty about whether assay really is in aqueous phase/uncertainty about EC number and reaction buffer
V197D Stability - Incubation Activity after incubation (1 hour at 30Β°C), measured by absorbance spectrophotometry (unsaturated bonds absorbance measurement, 235 nm) Ethanol 5% 100 124 123 % Incubation: 50 mM Tris-HCl buffer, Assay: 50 mM Tris-HCl buffer, 0.3 MM CaCl2 Incubation: 8, Assay: 8 Incubation: 30Β°C, Assay: 30Β°C 0.5% Polygalacturonic acid Ξ”4,5-unsaturated oligogalacturonic acids 0.3 mM Ca²⁺ β€” Water-incubated control (in %), assay in aqueous phase | Uncertainty about whether assay really is in aqueous phase/uncertainty about EC number and reaction buffer
I140V Stability - Incubation Activity after incubation (1 hour at 30Β°C), measured by absorbance spectrophotometry (unsaturated bonds absorbance measurement, 235 nm) Acetone 5% 100 105 106 % Incubation: 50 mM Tris-HCl buffer, Assay: 50 mM Tris-HCl buffer, 0.3 MM CaCl2 Incubation: 8, Assay: 8 Incubation: 30Β°C, Assay: 30Β°C 0.5% Polygalacturonic acid Ξ”4,5-unsaturated oligogalacturonic acids 0.3 mM Ca²⁺ β€” Water-incubated control (in %), assay in aqueous phase | Uncertainty about whether assay really is in aqueous phase/uncertainty about EC number and reaction buffer
V197D Stability - Incubation Activity after incubation (1 hour at 30Β°C), measured by absorbance spectrophotometry (unsaturated bonds absorbance measurement, 235 nm) Acetone 5% 100 105 110 % Incubation: 50 mM Tris-HCl buffer, Assay: 50 mM Tris-HCl buffer, 0.3 MM CaCl2 Incubation: 8, Assay: 8 Incubation: 30Β°C, Assay: 30Β°C 0.5% Polygalacturonic acid Ξ”4,5-unsaturated oligogalacturonic acids 0.3 mM Ca²⁺ β€” Water-incubated control (in %), assay in aqueous phase | Uncertainty about whether assay really is in aqueous phase/uncertainty about EC number and reaction buffer
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Mutations in this dataset (4)

WT I140V I140V/V197D V197D

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.

Structure

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