Fumarate hydratase class II from Thermus thermophilus HB8

Enzyme Description

Species
Thermus thermophilus HB8 β†—
Extremophile
Yes "thermophilic" organism
EC Number
4.2.1.2 β†—

Sequence

Length: 466 amino acids
MEYRIERDTMGEVRVPADKYWGAQTQRSLENFRIGTDRFRMPLEIIRAYGMLKKAAARANLELGELPEEIAKAIIQAAEEVVQGKWDDHFPLVVFQTGSGTQTNMNVNEVIANRASEILGKPLGSKYVHPNDHVNRGQSSNDTFPTAMYVAVALALHQRLYPAVEGLIRTFTAKAQAFDQIVKVGRTHLMDAVPITLGQEIGSWAAQLKTTLAAVKEMEKGLYNLAIGGTAVGTGLNAHPRFGELVAKYLAEETGLPFRVAENRFAALAAHDELVNVMGAIRTLAGALMKIGNDVRWLASGPYAGIGEITIPANEPGSSIMPGKVNPTQVEALTMVVVRVYGNDHTVAFAGSQGNFQLNVYKPVMAYSTLESINLLADAVASFDAHLAQGIEPNLERIEEHLQKNPMLATALNKAIGYDKAAEIVKKALKEKKTLKQAALELGYLTEEEFDRIVVPMRLAKPHEGA
Yanhui Liu et al. (2015) β€” Production of Fumaric Acid from l-Malic Acid by Solvent Engineering Using a Recombinant Thermostable Fumarase from Thermus thermophilus HB8
Applied Biochemistry and Biotechnology  Β· doi:10.1007/s12010-014-1468-z β†—  Β· Stability - Incubation Activity + Stability - Conversion
64 measurements ▼
Database ID
UniProt: Q5SKT5 β†—
Sequence Annotation
Inferred - from protein name
Protein Source
Recombinant, host bacterium Escherichia coli BL21 (DE3)

Experimental Data (64 measurements)

64 measurements β€” page 3 of 4
Property Assay Solvent Solvent Volume Aqueous Reference Measured Value Units Solution pH Temperature Substrate(s) Product(s) Cofactor(s) Shaking Comments
Activity + Stability - Conversion Conversion after incubation (30 minutes at 70Β°C), measured by absorbance spectrophotometry (fumaric acid absorbance measurment, 250 nm) in the presence of organic solvent Ethylene Glycol 30% 17.1 20.2 % 100 mM Tris– HCl buffer 7.8 70Β°C 100 mM L-Malic acid Fumarate β€” β€” Classical aqueous control (in %)
Activity + Stability - Conversion Conversion after incubation (120 minutes at 70Β°C), measured by absorbance spectrophotometry (fumaric acid absorbance measurment, 250 nm) in the presence of organic solvent Ethylene Glycol 30% 19.1 25.1 % 100 mM Tris– HCl buffer 7.8 70Β°C 100 mM L-Malic acid Fumarate β€” β€” Classical aqueous control (in %)
Activity + Stability - Conversion Conversion after incubation (120 minutes at 70Β°C), measured by absorbance spectrophotometry (fumaric acid absorbance measurment, 250 nm) in the presence of organic solvent Methanol 70% 19.1 2.7 % 100 mM Tris– HCl buffer 7.8 70Β°C 100 mM L-Malic acid Fumarate β€” β€” Classical aqueous control (in %)
Activity + Stability - Conversion Conversion after incubation (30 minutes at 70Β°C), measured by absorbance spectrophotometry (fumaric acid absorbance measurment, 250 nm) in the presence of organic solvent Methanol 70% 17.1 2.1 % 100 mM Tris– HCl buffer 7.8 70Β°C 100 mM L-Malic acid Fumarate β€” β€” Classical aqueous control (in %)
Activity + Stability - Conversion Conversion after incubation (10 minutes at 70Β°C), measured by absorbance spectrophotometry (fumaric acid absorbance measurment, 250 nm) in the presence of organic solvent Methanol 70% 15.9 5.0 % 100 mM Tris– HCl buffer 7.8 70Β°C 100 mM L-Malic acid Fumarate β€” β€” Classical aqueous control (in %)
Activity + Stability - Conversion Conversion after incubation (360 minutes at 70Β°C), measured by absorbance spectrophotometry (fumaric acid absorbance measurment, 250 nm) in the presence of organic solvent Methanol 50% 24.0 8.6 % 100 mM Tris– HCl buffer 7.8 70Β°C 100 mM L-Malic acid Fumarate β€” β€” Classical aqueous control (in %)
Activity + Stability - Conversion Conversion after incubation (240 minutes at 70Β°C), measured by absorbance spectrophotometry (fumaric acid absorbance measurment, 250 nm) in the presence of organic solvent Methanol 50% 23.3 8.6 % 100 mM Tris– HCl buffer 7.8 70Β°C 100 mM L-Malic acid Fumarate β€” β€” Classical aqueous control (in %)
Activity + Stability - Conversion Conversion after incubation (120 minutes at 70Β°C), measured by absorbance spectrophotometry (fumaric acid absorbance measurment, 250 nm) in the presence of organic solvent Methanol 50% 19.1 9.1 % 100 mM Tris– HCl buffer 7.8 70Β°C 100 mM L-Malic acid Fumarate β€” β€” Classical aqueous control (in %)
Activity + Stability - Conversion Conversion after incubation (60 minutes at 70Β°C), measured by absorbance spectrophotometry (fumaric acid absorbance measurment, 250 nm) in the presence of organic solvent Methanol 50% 17.3 8.4 % 100 mM Tris– HCl buffer 7.8 70Β°C 100 mM L-Malic acid Fumarate β€” β€” Classical aqueous control (in %)
Activity + Stability - Conversion Conversion after incubation (30 minutes at 70Β°C), measured by absorbance spectrophotometry (fumaric acid absorbance measurment, 250 nm) in the presence of organic solvent Methanol 50% 17.1 7.9 % 100 mM Tris– HCl buffer 7.8 70Β°C 100 mM L-Malic acid Fumarate β€” β€” Classical aqueous control (in %)
Activity + Stability - Conversion Conversion after incubation (10 minutes at 70Β°C), measured by absorbance spectrophotometry (fumaric acid absorbance measurment, 250 nm) in the presence of organic solvent Methanol 50% 15.9 9.2 % 100 mM Tris– HCl buffer 7.8 70Β°C 100 mM L-Malic acid Fumarate β€” β€” Classical aqueous control (in %)
Activity + Stability - Conversion Conversion after incubation (360 minutes at 70Β°C), measured by absorbance spectrophotometry (fumaric acid absorbance measurment, 250 nm) in the presence of organic solvent Ethylene Glycol 30% 24.0 32.3 % 100 mM Tris– HCl buffer 7.8 70Β°C 100 mM L-Malic acid Fumarate β€” β€” Classical aqueous control (in %)
Activity + Stability - Conversion Conversion after incubation (240 minutes at 70Β°C), measured by absorbance spectrophotometry (fumaric acid absorbance measurment, 250 nm) in the presence of organic solvent Ethylene Glycol 30% 23.3 32.3 % 100 mM Tris– HCl buffer 7.8 70Β°C 100 mM L-Malic acid Fumarate β€” β€” Classical aqueous control (in %)
Activity + Stability - Conversion Conversion after incubation (60 minutes at 70Β°C), measured by absorbance spectrophotometry (fumaric acid absorbance measurment, 250 nm) in the presence of organic solvent Ethylene Glycol 30% 17.3 22.3 % 100 mM Tris– HCl buffer 7.8 70Β°C 100 mM L-Malic acid Fumarate β€” β€” Classical aqueous control (in %)
Stability - Incubation Activity after incubation (1 hour at 70Β°C), measured by absorbance spectrophotometry (fumaric acid absorbance measurment, 250 nm) in aqueous phase Ethylene Glycol 50% 100.0 106.7 % Incubation: 100 mM Tris– HCl buffer, Assay: 100 mM Tris– HCl buffer Incubation: 7.8, Assay: 7.8 Incubation: 70Β°C, Assay: 70Β°C 100 mM L-Malic acid Fumarate β€” β€” Non-incubated control (in %), assay measured in aqueous phase
Stability - Incubation Activity after incubation (3 hours at 70Β°C), measured by absorbance spectrophotometry (fumaric acid absorbance measurment, 250 nm) in aqueous phase Ethylene Glycol 50% 100.0 112.5 % Incubation: 100 mM Tris– HCl buffer, Assay: 100 mM Tris– HCl buffer Incubation: 7.8, Assay: 7.8 Incubation: 70Β°C, Assay: 70Β°C 100 mM L-Malic acid Fumarate β€” β€” Non-incubated control (in %), assay measured in aqueous phase
Stability - Incubation Activity after incubation (8 hours at 70Β°C), measured by absorbance spectrophotometry (fumaric acid absorbance measurment, 250 nm) in aqueous phase Ethylene Glycol 50% 100.0 119.6 % Incubation: 100 mM Tris– HCl buffer, Assay: 100 mM Tris– HCl buffer Incubation: 7.8, Assay: 7.8 Incubation: 70Β°C, Assay: 70Β°C 100 mM L-Malic acid Fumarate β€” β€” Non-incubated control (in %), assay measured in aqueous phase
Stability - Incubation Activity after incubation (30 hours at 70Β°C), measured by absorbance spectrophotometry (fumaric acid absorbance measurment, 250 nm) in aqueous phase Ethylene Glycol 50% 100.0 108.0 % Incubation: 100 mM Tris– HCl buffer, Assay: 100 mM Tris– HCl buffer Incubation: 7.8, Assay: 7.8 Incubation: 70Β°C, Assay: 70Β°C 100 mM L-Malic acid Fumarate β€” β€” Non-incubated control (in %), assay measured in aqueous phase
Stability - Incubation Activity after incubation (48 hours at 70Β°C), measured by absorbance spectrophotometry (fumaric acid absorbance measurment, 250 nm) in aqueous phase Ethylene Glycol 50% 100.0 104.0 % Incubation: 100 mM Tris– HCl buffer, Assay: 100 mM Tris– HCl buffer Incubation: 7.8, Assay: 7.8 Incubation: 70Β°C, Assay: 70Β°C 100 mM L-Malic acid Fumarate β€” β€” Non-incubated control (in %), assay measured in aqueous phase
Stability - Incubation Activity after incubation (1 hour at 70Β°C), measured by absorbance spectrophotometry (fumaric acid absorbance measurment, 250 nm) in aqueous phase Ethylene Glycol 70% 100.0 98.2 % Incubation: 100 mM Tris– HCl buffer, Assay: 100 mM Tris– HCl buffer Incubation: 7.8, Assay: 7.8 Incubation: 70Β°C, Assay: 70Β°C 100 mM L-Malic acid Fumarate β€” β€” Non-incubated control (in %), assay measured in aqueous phase
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Visualization : Stability β€” Incubation

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

Visualization : Activity + Stability β€” Conversion

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

Structure

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