Fumarate hydratase class II from Thermus thermophilus HB8

Enzyme Description

Species

Thermus thermophilus HB8 ↗

Extremophile

Yes "thermophilic" organism

EC Number

4.2.1.2 ↗

Sequence

Length: 466 amino acids

MEYRIERDTMGEVRVPADKYWGAQTQRSLENFRIGTDRFRMPLEIIRAYGMLKKAAARANLELGELPEEIAKAIIQAAEEVVQGKWDDHFPLVVFQTGSGTQTNMNVNEVIANRASEILGKPLGSKYVHPNDHVNRGQSSNDTFPTAMYVAVALALHQRLYPAVEGLIRTFTAKAQAFDQIVKVGRTHLMDAVPITLGQEIGSWAAQLKTTLAAVKEMEKGLYNLAIGGTAVGTGLNAHPRFGELVAKYLAEETGLPFRVAENRFAALAAHDELVNVMGAIRTLAGALMKIGNDVRWLASGPYAGIGEITIPANEPGSSIMPGKVNPTQVEALTMVVVRVYGNDHTVAFAGSQGNFQLNVYKPVMAYSTLESINLLADAVASFDAHLAQGIEPNLERIEEHLQKNPMLATALNKAIGYDKAAEIVKKALKEKKTLKQAALELGYLTEEEFDRIVVPMRLAKPHEGA

Yanhui Liu et al. (2015) — Production of Fumaric Acid from l-Malic Acid by Solvent Engineering Using a Recombinant Thermostable Fumarase from Thermus thermophilus HB8

Applied Biochemistry and Biotechnology · doi:10.1007/s12010-014-1468-z ↗ · Stability - Incubation Activity + Stability - Conversion

64 measurements ▼

Database ID

UniProt: Q5SKT5 ↗

Sequence Annotation

Inferred - from protein name

Protein Source

Recombinant, host bacterium Escherichia coli BL21 (DE3)

Experimental Data (64 measurements)

64 measurements — page 4 of 4

Property	Assay	Solvent	Solvent Volume	Aqueous Reference	Measured Value	Units	Solution	pH	Temperature	Substrate(s)	Product(s)	Cofactor(s)	Shaking	Comments
Stability - Incubation	Activity after incubation (3 hours at 70°C), measured by absorbance spectrophotometry (fumaric acid absorbance measurment, 250 nm) in aqueous phase	Ethylene Glycol	70%	100.0	104.0	%	Incubation: 100 mM Tris– HCl buffer, Assay: 100 mM Tris– HCl buffer	Incubation: 7.8, Assay: 7.8	Incubation: 70°C, Assay: 70°C	100 mM L-Malic acid	Fumarate	—	—	Non-incubated control (in %), assay measured in aqueous phase
Stability - Incubation	Activity after incubation (8 hours at 70°C), measured by absorbance spectrophotometry (fumaric acid absorbance measurment, 250 nm) in aqueous phase	Ethylene Glycol	70%	100.0	96.0	%	Incubation: 100 mM Tris– HCl buffer, Assay: 100 mM Tris– HCl buffer	Incubation: 7.8, Assay: 7.8	Incubation: 70°C, Assay: 70°C	100 mM L-Malic acid	Fumarate	—	—	Non-incubated control (in %), assay measured in aqueous phase
Stability - Incubation	Activity after incubation (30 hours at 70°C), measured by absorbance spectrophotometry (fumaric acid absorbance measurment, 250 nm) in aqueous phase	Ethylene Glycol	70%	100.0	76.3	%	Incubation: 100 mM Tris– HCl buffer, Assay: 100 mM Tris– HCl buffer	Incubation: 7.8, Assay: 7.8	Incubation: 70°C, Assay: 70°C	100 mM L-Malic acid	Fumarate	—	—	Non-incubated control (in %), assay measured in aqueous phase
Stability - Incubation	Activity after incubation (48 hours at 70°C), measured by absorbance spectrophotometry (fumaric acid absorbance measurment, 250 nm) in aqueous phase	Ethylene Glycol	70%	100.0	77.7	%	Incubation: 100 mM Tris– HCl buffer, Assay: 100 mM Tris– HCl buffer	Incubation: 7.8, Assay: 7.8	Incubation: 70°C, Assay: 70°C	100 mM L-Malic acid	Fumarate	—	—	Non-incubated control (in %), assay measured in aqueous phase

Activity + Stability - Conversion	Conversion after incubation (360 minutes at 70°C), measured by absorbance spectrophotometry (fumaric acid absorbance measurment, 250 nm) in the presence of organic solvent	Ethylene Glycol	50%	24.0	36.8	%	100 mM Tris– HCl buffer	7.8	70°C	100 mM L-Malic acid	Fumarate	—	—	Classical aqueous control (in %)
Activity + Stability - Conversion	Conversion after incubation (240 minutes at 70°C), measured by absorbance spectrophotometry (fumaric acid absorbance measurment, 250 nm) in the presence of organic solvent	Methanol	70%	23.3	2.0	%	100 mM Tris– HCl buffer	7.8	70°C	100 mM L-Malic acid	Fumarate	—	—	Classical aqueous control (in %)
Activity + Stability - Conversion	Conversion after incubation (360 minutes at 70°C), measured by absorbance spectrophotometry (fumaric acid absorbance measurment, 250 nm) in the presence of organic solvent	Methanol	70%	24.0	2.6	%	100 mM Tris– HCl buffer	7.8	70°C	100 mM L-Malic acid	Fumarate	—	—	Classical aqueous control (in %)
Activity + Stability - Conversion	Conversion after incubation (10 minutes at 70°C), measured by absorbance spectrophotometry (fumaric acid absorbance measurment, 250 nm) in the presence of organic solvent	Methanol	90%	15.9	1.1	%	100 mM Tris– HCl buffer	7.8	70°C	100 mM L-Malic acid	Fumarate	—	—	Classical aqueous control (in %)
Activity + Stability - Conversion	Conversion after incubation (30 minutes at 70°C), measured by absorbance spectrophotometry (fumaric acid absorbance measurment, 250 nm) in the presence of organic solvent	Methanol	90%	17.1	1.3	%	100 mM Tris– HCl buffer	7.8	70°C	100 mM L-Malic acid	Fumarate	—	—	Classical aqueous control (in %)
Activity + Stability - Conversion	Conversion after incubation (60 minutes at 70°C), measured by absorbance spectrophotometry (fumaric acid absorbance measurment, 250 nm) in the presence of organic solvent	Methanol	90%	17.3	1.8	%	100 mM Tris– HCl buffer	7.8	70°C	100 mM L-Malic acid	Fumarate	—	—	Classical aqueous control (in %)
Activity + Stability - Conversion	Conversion after incubation (120 minutes at 70°C), measured by absorbance spectrophotometry (fumaric acid absorbance measurment, 250 nm) in the presence of organic solvent	Methanol	90%	19.1	1.1	%	100 mM Tris– HCl buffer	7.8	70°C	100 mM L-Malic acid	Fumarate	—	—	Classical aqueous control (in %)
Activity + Stability - Conversion	Conversion after incubation (240 minutes at 70°C), measured by absorbance spectrophotometry (fumaric acid absorbance measurment, 250 nm) in the presence of organic solvent	Methanol	90%	23.3	0.2	%	100 mM Tris– HCl buffer	7.8	70°C	100 mM L-Malic acid	Fumarate	—	—	Classical aqueous control (in %)
Activity + Stability - Conversion	Conversion after incubation (360 minutes at 70°C), measured by absorbance spectrophotometry (fumaric acid absorbance measurment, 250 nm) in the presence of organic solvent	Methanol	90%	24.0	1.5	%	100 mM Tris– HCl buffer	7.8	70°C	100 mM L-Malic acid	Fumarate	—	—	Classical aqueous control (in %)
Activity + Stability - Conversion	Conversion after incubation (10 minutes at 70°C), measured by absorbance spectrophotometry (fumaric acid absorbance measurment, 250 nm) in the presence of organic solvent	Ethylene Glycol	50%	15.9	16.9	%	100 mM Tris– HCl buffer	7.8	70°C	100 mM L-Malic acid	Fumarate	—	—	Classical aqueous control (in %)
Activity + Stability - Conversion	Conversion after incubation (30 minutes at 70°C), measured by absorbance spectrophotometry (fumaric acid absorbance measurment, 250 nm) in the presence of organic solvent	Ethylene Glycol	50%	17.1	20.7	%	100 mM Tris– HCl buffer	7.8	70°C	100 mM L-Malic acid	Fumarate	—	—	Classical aqueous control (in %)
Activity + Stability - Conversion	Conversion after incubation (60 minutes at 70°C), measured by absorbance spectrophotometry (fumaric acid absorbance measurment, 250 nm) in the presence of organic solvent	Ethylene Glycol	50%	17.3	22.1	%	100 mM Tris– HCl buffer	7.8	70°C	100 mM L-Malic acid	Fumarate	—	—	Classical aqueous control (in %)
Activity + Stability - Conversion	Conversion after incubation (120 minutes at 70°C), measured by absorbance spectrophotometry (fumaric acid absorbance measurment, 250 nm) in the presence of organic solvent	Ethylene Glycol	50%	19.1	26.7	%	100 mM Tris– HCl buffer	7.8	70°C	100 mM L-Malic acid	Fumarate	—	—	Classical aqueous control (in %)
Activity + Stability - Conversion	Conversion after incubation (240 minutes at 70°C), measured by absorbance spectrophotometry (fumaric acid absorbance measurment, 250 nm) in the presence of organic solvent	Ethylene Glycol	50%	23.3	38.4	%	100 mM Tris– HCl buffer	7.8	70°C	100 mM L-Malic acid	Fumarate	—	—	Classical aqueous control (in %)
Activity + Stability - Conversion	Conversion after incubation (30 minutes at 70°C), measured by absorbance spectrophotometry (fumaric acid absorbance measurment, 250 nm) in the presence of organic solvent	Methanol	30%	17.1	21.7	%	100 mM Tris– HCl buffer	7.8	70°C	100 mM L-Malic acid	Fumarate	—	—	Classical aqueous control (in %)
Activity + Stability - Conversion	Conversion after incubation (10 minutes at 70°C), measured by absorbance spectrophotometry (fumaric acid absorbance measurment, 250 nm) in the presence of organic solvent	Ethylene Glycol	70%	15.9	18.7	%	100 mM Tris– HCl buffer	7.8	70°C	100 mM L-Malic acid	Fumarate	—	—	Classical aqueous control (in %)
Activity + Stability - Conversion	Conversion after incubation (30 minutes at 70°C), measured by absorbance spectrophotometry (fumaric acid absorbance measurment, 250 nm) in the presence of organic solvent	Ethylene Glycol	70%	17.1	25.2	%	100 mM Tris– HCl buffer	7.8	70°C	100 mM L-Malic acid	Fumarate	—	—	Classical aqueous control (in %)
Activity + Stability - Conversion	Conversion after incubation (60 minutes at 70°C), measured by absorbance spectrophotometry (fumaric acid absorbance measurment, 250 nm) in the presence of organic solvent	Ethylene Glycol	70%	17.3	26.3	%	100 mM Tris– HCl buffer	7.8	70°C	100 mM L-Malic acid	Fumarate	—	—	Classical aqueous control (in %)
Activity + Stability - Conversion	Conversion after incubation (120 minutes at 70°C), measured by absorbance spectrophotometry (fumaric acid absorbance measurment, 250 nm) in the presence of organic solvent	Ethylene Glycol	70%	19.1	32.4	%	100 mM Tris– HCl buffer	7.8	70°C	100 mM L-Malic acid	Fumarate	—	—	Classical aqueous control (in %)
Activity + Stability - Conversion	Conversion after incubation (240 minutes at 70°C), measured by absorbance spectrophotometry (fumaric acid absorbance measurment, 250 nm) in the presence of organic solvent	Ethylene Glycol	70%	23.3	46.3	%	100 mM Tris– HCl buffer	7.8	70°C	100 mM L-Malic acid	Fumarate	—	—	Classical aqueous control (in %)
Activity + Stability - Conversion	Conversion after incubation (360 minutes at 70°C), measured by absorbance spectrophotometry (fumaric acid absorbance measurment, 250 nm) in the presence of organic solvent	Ethylene Glycol	70%	24.0	45.5	%	100 mM Tris– HCl buffer	7.8	70°C	100 mM L-Malic acid	Fumarate	—	—	Classical aqueous control (in %)
Activity + Stability - Conversion	Conversion after incubation (10 minutes at 70°C), measured by absorbance spectrophotometry (fumaric acid absorbance measurment, 250 nm) in the presence of organic solvent	Ethylene Glycol	90%	15.9	3.7	%	100 mM Tris– HCl buffer	7.8	70°C	100 mM L-Malic acid	Fumarate	—	—	Classical aqueous control (in %)
Activity + Stability - Conversion	Conversion after incubation (30 minutes at 70°C), measured by absorbance spectrophotometry (fumaric acid absorbance measurment, 250 nm) in the presence of organic solvent	Ethylene Glycol	90%	17.1	3.5	%	100 mM Tris– HCl buffer	7.8	70°C	100 mM L-Malic acid	Fumarate	—	—	Classical aqueous control (in %)
Activity + Stability - Conversion	Conversion after incubation (60 minutes at 70°C), measured by absorbance spectrophotometry (fumaric acid absorbance measurment, 250 nm) in the presence of organic solvent	Ethylene Glycol	90%	17.3	2.5	%	100 mM Tris– HCl buffer	7.8	70°C	100 mM L-Malic acid	Fumarate	—	—	Classical aqueous control (in %)
Activity + Stability - Conversion	Conversion after incubation (120 minutes at 70°C), measured by absorbance spectrophotometry (fumaric acid absorbance measurment, 250 nm) in the presence of organic solvent	Ethylene Glycol	90%	19.1	1.1	%	100 mM Tris– HCl buffer	7.8	70°C	100 mM L-Malic acid	Fumarate	—	—	Classical aqueous control (in %)
Activity + Stability - Conversion	Conversion after incubation (240 minutes at 70°C), measured by absorbance spectrophotometry (fumaric acid absorbance measurment, 250 nm) in the presence of organic solvent	Ethylene Glycol	90%	23.3	0.2	%	100 mM Tris– HCl buffer	7.8	70°C	100 mM L-Malic acid	Fumarate	—	—	Classical aqueous control (in %)
Activity + Stability - Conversion	Conversion after incubation (360 minutes at 70°C), measured by absorbance spectrophotometry (fumaric acid absorbance measurment, 250 nm) in the presence of organic solvent	Ethylene Glycol	90%	24.0	1.5	%	100 mM Tris– HCl buffer	7.8	70°C	100 mM L-Malic acid	Fumarate	—	—	Classical aqueous control (in %)
Activity + Stability - Conversion	Conversion after incubation (60 minutes at 70°C), measured by absorbance spectrophotometry (fumaric acid absorbance measurment, 250 nm) in the presence of organic solvent	Methanol	70%	17.3	1.8	%	100 mM Tris– HCl buffer	7.8	70°C	100 mM L-Malic acid	Fumarate	—	—	Classical aqueous control (in %)
Activity + Stability - Conversion	Conversion after incubation (10 minutes at 70°C), measured by absorbance spectrophotometry (fumaric acid absorbance measurment, 250 nm) in the presence of organic solvent	Methanol	30%	15.9	21.8	%	100 mM Tris– HCl buffer	7.8	70°C	100 mM L-Malic acid	Fumarate	—	—	Classical aqueous control (in %)
Activity + Stability - Conversion	Conversion after incubation (60 minutes at 70°C), measured by absorbance spectrophotometry (fumaric acid absorbance measurment, 250 nm) in the presence of organic solvent	Methanol	30%	17.3	23.8	%	100 mM Tris– HCl buffer	7.8	70°C	100 mM L-Malic acid	Fumarate	—	—	Classical aqueous control (in %)
Activity + Stability - Conversion	Conversion after incubation (120 minutes at 70°C), measured by absorbance spectrophotometry (fumaric acid absorbance measurment, 250 nm) in the presence of organic solvent	Methanol	30%	19.1	27.6	%	100 mM Tris– HCl buffer	7.8	70°C	100 mM L-Malic acid	Fumarate	—	—	Classical aqueous control (in %)
Activity + Stability - Conversion	Conversion after incubation (240 minutes at 70°C), measured by absorbance spectrophotometry (fumaric acid absorbance measurment, 250 nm) in the presence of organic solvent	Methanol	30%	23.3	35.5	%	100 mM Tris– HCl buffer	7.8	70°C	100 mM L-Malic acid	Fumarate	—	—	Classical aqueous control (in %)
Activity + Stability - Conversion	Conversion after incubation (360 minutes at 70°C), measured by absorbance spectrophotometry (fumaric acid absorbance measurment, 250 nm) in the presence of organic solvent	Methanol	30%	24.0	35.0	%	100 mM Tris– HCl buffer	7.8	70°C	100 mM L-Malic acid	Fumarate	—	—	Classical aqueous control (in %)
Activity + Stability - Conversion	Conversion after incubation (10 minutes at 70°C), measured by absorbance spectrophotometry (fumaric acid absorbance measurment, 250 nm) in the presence of organic solvent	tert-Butanol	30%	15.9	3.0	%	100 mM Tris– HCl buffer	7.8	70°C	100 mM L-Malic acid	Fumarate	—	—	Classical aqueous control (in %)
Activity + Stability - Conversion	Conversion after incubation (30 minutes at 70°C), measured by absorbance spectrophotometry (fumaric acid absorbance measurment, 250 nm) in the presence of organic solvent	tert-Butanol	30%	17.1	3.0	%	100 mM Tris– HCl buffer	7.8	70°C	100 mM L-Malic acid	Fumarate	—	—	Classical aqueous control (in %)
Activity + Stability - Conversion	Conversion after incubation (60 minutes at 70°C), measured by absorbance spectrophotometry (fumaric acid absorbance measurment, 250 nm) in the presence of organic solvent	tert-Butanol	30%	17.3	4.5	%	100 mM Tris– HCl buffer	7.8	70°C	100 mM L-Malic acid	Fumarate	—	—	Classical aqueous control (in %)
Activity + Stability - Conversion	Conversion after incubation (120 minutes at 70°C), measured by absorbance spectrophotometry (fumaric acid absorbance measurment, 250 nm) in the presence of organic solvent	tert-Butanol	30%	19.1	7.5	%	100 mM Tris– HCl buffer	7.8	70°C	100 mM L-Malic acid	Fumarate	—	—	Classical aqueous control (in %)
Activity + Stability - Conversion	Conversion after incubation (240 minutes at 70°C), measured by absorbance spectrophotometry (fumaric acid absorbance measurment, 250 nm) in the presence of organic solvent	tert-Butanol	30%	23.3	2.9	%	100 mM Tris– HCl buffer	7.8	70°C	100 mM L-Malic acid	Fumarate	—	—	Classical aqueous control (in %)
Activity + Stability - Conversion	Conversion after incubation (360 minutes at 70°C), measured by absorbance spectrophotometry (fumaric acid absorbance measurment, 250 nm) in the presence of organic solvent	tert-Butanol	30%	24.0	3.1	%	100 mM Tris– HCl buffer	7.8	70°C	100 mM L-Malic acid	Fumarate	—	—	Classical aqueous control (in %)
Activity + Stability - Conversion	Conversion after incubation (10 minutes at 70°C), measured by absorbance spectrophotometry (fumaric acid absorbance measurment, 250 nm) in the presence of organic solvent	Ethylene Glycol	30%	15.9	17.8	%	100 mM Tris– HCl buffer	7.8	70°C	100 mM L-Malic acid	Fumarate	—	—	Classical aqueous control (in %)
Activity + Stability - Conversion	Conversion after incubation (30 minutes at 70°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	70°C	100 mM L-Malic acid	Fumarate	—	—	Classical aqueous control (in %)
Activity + Stability - Conversion	Conversion after incubation (120 minutes at 70°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	70°C	100 mM L-Malic acid	Fumarate	—	—	Classical aqueous control (in %)
Activity + Stability - Conversion	Conversion after incubation (120 minutes at 70°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	70°C	100 mM L-Malic acid	Fumarate	—	—	Classical aqueous control (in %)
Activity + Stability - Conversion	Conversion after incubation (30 minutes at 70°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	70°C	100 mM L-Malic acid	Fumarate	—	—	Classical aqueous control (in %)
Activity + Stability - Conversion	Conversion after incubation (10 minutes at 70°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	70°C	100 mM L-Malic acid	Fumarate	—	—	Classical aqueous control (in %)
Activity + Stability - Conversion	Conversion after incubation (360 minutes at 70°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	70°C	100 mM L-Malic acid	Fumarate	—	—	Classical aqueous control (in %)
Activity + Stability - Conversion	Conversion after incubation (240 minutes at 70°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	70°C	100 mM L-Malic acid	Fumarate	—	—	Classical aqueous control (in %)
Activity + Stability - Conversion	Conversion after incubation (120 minutes at 70°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	70°C	100 mM L-Malic acid	Fumarate	—	—	Classical aqueous control (in %)
Activity + Stability - Conversion	Conversion after incubation (60 minutes at 70°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	70°C	100 mM L-Malic acid	Fumarate	—	—	Classical aqueous control (in %)
Activity + Stability - Conversion	Conversion after incubation (30 minutes at 70°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	70°C	100 mM L-Malic acid	Fumarate	—	—	Classical aqueous control (in %)
Activity + Stability - Conversion	Conversion after incubation (10 minutes at 70°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	70°C	100 mM L-Malic acid	Fumarate	—	—	Classical aqueous control (in %)
Activity + Stability - Conversion	Conversion after incubation (360 minutes at 70°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	70°C	100 mM L-Malic acid	Fumarate	—	—	Classical aqueous control (in %)
Activity + Stability - Conversion	Conversion after incubation (240 minutes at 70°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	70°C	100 mM L-Malic acid	Fumarate	—	—	Classical aqueous control (in %)
Activity + Stability - Conversion	Conversion after incubation (60 minutes at 70°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	70°C	100 mM L-Malic acid	Fumarate	—	—	Classical aqueous control (in %)
Stability - Incubation	Activity after incubation (1 hour at 70°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: 70°C, Assay: 70°C	100 mM L-Malic acid	Fumarate	—	—	Non-incubated control (in %), assay measured in aqueous phase
Stability - Incubation	Activity after incubation (3 hours at 70°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: 70°C, Assay: 70°C	100 mM L-Malic acid	Fumarate	—	—	Non-incubated control (in %), assay measured in aqueous phase
Stability - Incubation	Activity after incubation (8 hours at 70°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: 70°C, Assay: 70°C	100 mM L-Malic acid	Fumarate	—	—	Non-incubated control (in %), assay measured in aqueous phase
Stability - Incubation	Activity after incubation (30 hours at 70°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: 70°C, Assay: 70°C	100 mM L-Malic acid	Fumarate	—	—	Non-incubated control (in %), assay measured in aqueous phase
Stability - Incubation	Activity after incubation (48 hours at 70°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: 70°C, Assay: 70°C	100 mM L-Malic acid	Fumarate	—	—	Non-incubated control (in %), assay measured in aqueous phase
Stability - Incubation	Activity after incubation (1 hour at 70°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: 70°C, Assay: 70°C	100 mM L-Malic acid	Fumarate	—	—	Non-incubated control (in %), assay measured in aqueous phase
Stability - Incubation	Activity after incubation (3 hours at 70°C), measured by absorbance spectrophotometry (fumaric acid absorbance measurment, 250 nm) in aqueous phase	Ethylene Glycol	70%	100.0	104.0	%	Incubation: 100 mM Tris– HCl buffer, Assay: 100 mM Tris– HCl buffer	Incubation: 7.8, Assay: 7.8	Incubation: 70°C, Assay: 70°C	100 mM L-Malic acid	Fumarate	—	—	Non-incubated control (in %), assay measured in aqueous phase
Stability - Incubation	Activity after incubation (8 hours at 70°C), measured by absorbance spectrophotometry (fumaric acid absorbance measurment, 250 nm) in aqueous phase	Ethylene Glycol	70%	100.0	96.0	%	Incubation: 100 mM Tris– HCl buffer, Assay: 100 mM Tris– HCl buffer	Incubation: 7.8, Assay: 7.8	Incubation: 70°C, Assay: 70°C	100 mM L-Malic acid	Fumarate	—	—	Non-incubated control (in %), assay measured in aqueous phase
Stability - Incubation	Activity after incubation (30 hours at 70°C), measured by absorbance spectrophotometry (fumaric acid absorbance measurment, 250 nm) in aqueous phase	Ethylene Glycol	70%	100.0	76.3	%	Incubation: 100 mM Tris– HCl buffer, Assay: 100 mM Tris– HCl buffer	Incubation: 7.8, Assay: 7.8	Incubation: 70°C, Assay: 70°C	100 mM L-Malic acid	Fumarate	—	—	Non-incubated control (in %), assay measured in aqueous phase
Stability - Incubation	Activity after incubation (48 hours at 70°C), measured by absorbance spectrophotometry (fumaric acid absorbance measurment, 250 nm) in aqueous phase	Ethylene Glycol	70%	100.0	77.7	%	Incubation: 100 mM Tris– HCl buffer, Assay: 100 mM Tris– HCl buffer	Incubation: 7.8, Assay: 7.8	Incubation: 70°C, Assay: 70°C	100 mM L-Malic acid	Fumarate	—	—	Non-incubated control (in %), assay measured in aqueous phase

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Visualization : Stability — Incubation

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Visualization : Activity + Stability — Conversion