NADP-dependent malic enzyme from Sulfolobus solfataricus

Enzyme Description

Species

Sulfolobus solfataricus ↗

Extremophile

Yes "extreme thermoacidophilic" organism

EC Number

1.1.1.40 ↗

Sequence

Length: 427 amino acids

MVDAVELSRKYEGKIEIYPKVPITSYNDFALIYTPGVAEVSKAIYKDPDKSFELTSRWNTIAVVTDGTRVLGLGNIGPTAAMPVMEGKSLLFKYLGGVDAIPLPINVKSADEFTNVVKALETSFGGINLEDIESPKCFYILEKLQSMLNIPVWHDDQQGTAAAILAGLINAMILTNKDPKESKVVFYGAGASNIAAARILAKYGFKYENIVMIDSKGPLYADREDVDHLMLHHKWKYELAIKTNKWEAKEIKDAFSGADAVVAASTPGPNVIKKEWISLMNKEAIVFALANPVPEIWPQDAKEAGAKIVATGRSDFPNQVNNSLIFPAIFRGVLDSRAKAITDEMTIAASEALAKYARDRGINENYIIPRMDEWEAFYEEAAAVASKAVELGLARVKKSREEFREIAKERIIRARKIMNLVISTWSP

Annamaria Guagliardi et al. (1989) — Stability and activity of a thermostable malic enzyme in denaturants and water-miscible organic solvents

European Journal of Biochemistry · doi:10.1111/j.1432-1033.1989.tb14891.x ↗ · Activity - Classical Stability - Incubation

71 measurements ▼

Database ID

UniProt: A0A7S9IJ09 ↗

Sequence Annotation

Inferred - from protein name

Protein Source

Purified, archaeon Sulfolobus solfataricus

Experimental Data (71 measurements)

71 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 (2 hours at 25°C), measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm) in aqueous phase	Ethanol	50%	100.0	64.3	%	Incubation: 20 mM Tris/HCl buffer, Assay: 50 mM ammonium sulfate, 0.1 MM MnCl2	Incubation: 8, Assay: 8	Incubation: 25°C, Assay: 60°C	1 mM L-Malic acid	Carbon Dioxide (CO2) , Pyruvate	Assay: 0.05 mM NADP+	—	Non-incubated control (in %), assay in aqueous phase \| Clear about assay conditions
Stability - Incubation	Activity after incubation (1 hour at 25°C), measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm) in aqueous phase	Ethanol	50%	100.0	76.4	%	Incubation: 20 mM Tris/HCl buffer, Assay: 50 mM ammonium sulfate, 0.1 MM MnCl2	Incubation: 8, Assay: 8	Incubation: 25°C, Assay: 60°C	1 mM L-Malic acid	Carbon Dioxide (CO2) , Pyruvate	Assay: 0.05 mM NADP+	—	Non-incubated control (in %), assay in aqueous phase \| Clear about assay conditions
Stability - Incubation	Activity after incubation (24 hours at 25°C), measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm) in aqueous phase	Isopropanol	30%	100.0	20.7	%	Incubation: 20 mM Tris/HCl buffer, Assay: 50 mM ammonium sulfate, 0.1 MM MnCl2	Incubation: 8, Assay: 8	Incubation: 25°C, Assay: 60°C	1 mM L-Malic acid	Carbon Dioxide (CO2) , Pyruvate	Assay: 0.05 mM NADP+	—	Non-incubated control (in %), assay in aqueous phase \| Clear about assay conditions
Stability - Incubation	Activity after incubation (1 hour at 25°C), measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm) in aqueous phase	Isopropanol	10%	100.0	108.2	%	Incubation: 20 mM Tris/HCl buffer, Assay: 50 mM ammonium sulfate, 0.1 MM MnCl2	Incubation: 8, Assay: 8	Incubation: 25°C, Assay: 60°C	1 mM L-Malic acid	Carbon Dioxide (CO2) , Pyruvate	Assay: 0.05 mM NADP+	—	Non-incubated control (in %), assay in aqueous phase \| Clear about assay conditions
Stability - Incubation	Activity after incubation (8 hours at 25°C), measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm) in aqueous phase	Isopropanol	30%	100.0	58.9	%	Incubation: 20 mM Tris/HCl buffer, Assay: 50 mM ammonium sulfate, 0.1 MM MnCl2	Incubation: 8, Assay: 8	Incubation: 25°C, Assay: 60°C	1 mM L-Malic acid	Carbon Dioxide (CO2) , Pyruvate	Assay: 0.05 mM NADP+	—	Non-incubated control (in %), assay in aqueous phase \| Clear about assay conditions
Stability - Incubation	Activity after incubation (4 hours at 25°C), measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm) in aqueous phase	Isopropanol	30%	100.0	66.4	%	Incubation: 20 mM Tris/HCl buffer, Assay: 50 mM ammonium sulfate, 0.1 MM MnCl2	Incubation: 8, Assay: 8	Incubation: 25°C, Assay: 60°C	1 mM L-Malic acid	Carbon Dioxide (CO2) , Pyruvate	Assay: 0.05 mM NADP+	—	Non-incubated control (in %), assay in aqueous phase \| Clear about assay conditions
Stability - Incubation	Activity after incubation (2 hours at 25°C), measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm) in aqueous phase	Isopropanol	30%	100.0	80.7	%	Incubation: 20 mM Tris/HCl buffer, Assay: 50 mM ammonium sulfate, 0.1 MM MnCl2	Incubation: 8, Assay: 8	Incubation: 25°C, Assay: 60°C	1 mM L-Malic acid	Carbon Dioxide (CO2) , Pyruvate	Assay: 0.05 mM NADP+	—	Non-incubated control (in %), assay in aqueous phase \| Clear about assay conditions
Stability - Incubation	Activity after incubation (24 hours at 25°C), measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm) in aqueous phase	Ethanol	15%	100.0	85.4	%	Incubation: 20 mM Tris/HCl buffer, Assay: 50 mM ammonium sulfate, 0.1 MM MnCl2	Incubation: 8, Assay: 8	Incubation: 25°C, Assay: 60°C	1 mM L-Malic acid	Carbon Dioxide (CO2) , Pyruvate	Assay: 0.05 mM NADP+	—	Non-incubated control (in %), assay in aqueous phase \| Clear about assay conditions
Stability - Incubation	Activity after incubation (20 hours at 25°C), measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm) in aqueous phase	Ethanol	15%	100.0	85.7	%	Incubation: 20 mM Tris/HCl buffer, Assay: 50 mM ammonium sulfate, 0.1 MM MnCl2	Incubation: 8, Assay: 8	Incubation: 25°C, Assay: 60°C	1 mM L-Malic acid	Carbon Dioxide (CO2) , Pyruvate	Assay: 0.05 mM NADP+	—	Non-incubated control (in %), assay in aqueous phase \| Clear about assay conditions
Stability - Incubation	Activity after incubation (6 hours at 25°C), measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm) in aqueous phase	Ethanol	15%	100.0	86.1	%	Incubation: 20 mM Tris/HCl buffer, Assay: 50 mM ammonium sulfate, 0.1 MM MnCl2	Incubation: 8, Assay: 8	Incubation: 25°C, Assay: 60°C	1 mM L-Malic acid	Carbon Dioxide (CO2) , Pyruvate	Assay: 0.05 mM NADP+	—	Non-incubated control (in %), assay in aqueous phase \| Clear about assay conditions
Stability - Incubation	Activity after incubation (4 hours at 25°C), measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm) in aqueous phase	Ethanol	15%	100.0	87.1	%	Incubation: 20 mM Tris/HCl buffer, Assay: 50 mM ammonium sulfate, 0.1 MM MnCl2	Incubation: 8, Assay: 8	Incubation: 25°C, Assay: 60°C	1 mM L-Malic acid	Carbon Dioxide (CO2) , Pyruvate	Assay: 0.05 mM NADP+	—	Non-incubated control (in %), assay in aqueous phase \| Clear about assay conditions

Activity - Classical

Activity measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm) in the presence of organic solvent

Methanol

50%

100.0

138.6

20 mM Tris/HCl, 50 mM ammonium sulfate, 0.1 MM MnCl2

60 °C

1 mM L-Malic acid

Carbon Dioxide (CO2), Pyruvate

0.05 mM NADP+

—

Classical aqueous control (in %)

Activity - Classical

Activity measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm) in the presence of organic solvent

Methanol

100.0

105.5

20 mM Tris/HCl, 50 mM ammonium sulfate, 0.1 MM MnCl2

25 °C

1 mM L-Malic acid

Carbon Dioxide (CO2), Pyruvate

0.05 mM NADP+

—

Classical aqueous control (in %)

Activity - Classical

Activity measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm) in the presence of organic solvent

Methanol

10%

100.0

126.8

20 mM Tris/HCl, 50 mM ammonium sulfate, 0.1 MM MnCl2

25 °C

1 mM L-Malic acid

Carbon Dioxide (CO2), Pyruvate

0.05 mM NADP+

—

Classical aqueous control (in %)

Activity - Classical

Activity measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm) in the presence of organic solvent

Methanol

30%

100.0

141.6

20 mM Tris/HCl, 50 mM ammonium sulfate, 0.1 MM MnCl2

25 °C

1 mM L-Malic acid

Carbon Dioxide (CO2), Pyruvate

0.05 mM NADP+

—

Classical aqueous control (in %)

Activity - Classical

Activity measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm) in the presence of organic solvent

Methanol

40%

100.0

159.7

20 mM Tris/HCl, 50 mM ammonium sulfate, 0.1 MM MnCl2

25 °C

1 mM L-Malic acid

Carbon Dioxide (CO2), Pyruvate

0.05 mM NADP+

—

Classical aqueous control (in %)

Activity - Classical

Activity measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm) in the presence of organic solvent

Methanol

50%

100.0

199.4

20 mM Tris/HCl, 50 mM ammonium sulfate, 0.1 MM MnCl2

25 °C

1 mM L-Malic acid

Carbon Dioxide (CO2), Pyruvate

0.05 mM NADP+

—

Classical aqueous control (in %)

Activity - Classical

Activity measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm) in the presence of organic solvent

Methanol

60%

100.0

209.3

20 mM Tris/HCl, 50 mM ammonium sulfate, 0.1 MM MnCl2

25 °C

1 mM L-Malic acid

Carbon Dioxide (CO2), Pyruvate

0.05 mM NADP+

—

Classical aqueous control (in %)

Activity - Classical

Activity measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm) in the presence of organic solvent

Methanol

70%

100.0

203.6

20 mM Tris/HCl, 50 mM ammonium sulfate, 0.1 MM MnCl2

25 °C

1 mM L-Malic acid

Carbon Dioxide (CO2), Pyruvate

0.05 mM NADP+

—

Classical aqueous control (in %)

Activity - Classical

Activity measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm) in the presence of organic solvent

Ethanol

10%

100.0

122.7

20 mM Tris/HCl, 50 mM ammonium sulfate, 0.1 MM MnCl2

25 °C

1 mM L-Malic acid

Carbon Dioxide (CO2), Pyruvate

0.05 mM NADP+

—

Classical aqueous control (in %)

Activity - Classical

Activity measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm) in the presence of organic solvent

Ethanol

30%

100.0

135.6

20 mM Tris/HCl, 50 mM ammonium sulfate, 0.1 MM MnCl2

25 °C

1 mM L-Malic acid

Carbon Dioxide (CO2), Pyruvate

0.05 mM NADP+

—

Classical aqueous control (in %)

Activity - Classical

Activity measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm) in the presence of organic solvent

Ethanol

40%

100.0

153.4

20 mM Tris/HCl, 50 mM ammonium sulfate, 0.1 MM MnCl2

25 °C

1 mM L-Malic acid

Carbon Dioxide (CO2), Pyruvate

0.05 mM NADP+

—

Classical aqueous control (in %)

Activity - Classical

Activity measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm) in the presence of organic solvent

Ethanol

50%

100.0

172.3

20 mM Tris/HCl, 50 mM ammonium sulfate, 0.1 MM MnCl2

25 °C

1 mM L-Malic acid

Carbon Dioxide (CO2), Pyruvate

0.05 mM NADP+

—

Classical aqueous control (in %)

Activity - Classical

Activity measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm) in the presence of organic solvent

Ethanol

60%

100.0

157.3

20 mM Tris/HCl, 50 mM ammonium sulfate, 0.1 MM MnCl2

25 °C

1 mM L-Malic acid

Carbon Dioxide (CO2), Pyruvate

0.05 mM NADP+

—

Classical aqueous control (in %)

Activity - Classical

Activity measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm) in the presence of organic solvent

Ethanol

70%

100.0

142.2

20 mM Tris/HCl, 50 mM ammonium sulfate, 0.1 MM MnCl2

25 °C

1 mM L-Malic acid

Carbon Dioxide (CO2), Pyruvate

0.05 mM NADP+

—

Classical aqueous control (in %)

Activity - Classical

Activity measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm) in the presence of organic solvent

Acetone

10%

100.0

120.0

20 mM Tris/HCl, 50 mM ammonium sulfate, 0.1 MM MnCl2

25 °C

1 mM L-Malic acid

Carbon Dioxide (CO2), Pyruvate

0.05 mM NADP+

—

Classical aqueous control (in %)

Activity - Classical

Activity measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm) in the presence of organic solvent

Acetone

30%

100.0

146.0

20 mM Tris/HCl, 50 mM ammonium sulfate, 0.1 MM MnCl2

25 °C

1 mM L-Malic acid

Carbon Dioxide (CO2), Pyruvate

0.05 mM NADP+

—

Classical aqueous control (in %)

Activity - Classical

Activity measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm) in the presence of organic solvent

Acetone

50%

100.0

133.4

20 mM Tris/HCl, 50 mM ammonium sulfate, 0.1 MM MnCl2

25 °C

1 mM L-Malic acid

Carbon Dioxide (CO2), Pyruvate

0.05 mM NADP+

—

Classical aqueous control (in %)

Activity - Classical

Activity measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm) in the presence of organic solvent

Methanol

30%

100.0

126.0

20 mM Tris/HCl, 50 mM ammonium sulfate, 0.1 MM MnCl2

60 °C

1 mM L-Malic acid

Carbon Dioxide (CO2), Pyruvate

0.05 mM NADP+

—

Classical aqueous control (in %)

Activity - Classical

Activity measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm) in the presence of organic solvent

Methanol

10%

100.0

113.7

20 mM Tris/HCl, 50 mM ammonium sulfate, 0.1 MM MnCl2

60 °C

1 mM L-Malic acid

Carbon Dioxide (CO2), Pyruvate

0.05 mM NADP+

—

Classical aqueous control (in %)

Activity - Classical

Activity measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm) in the presence of organic solvent

Methanol

50%

100.0

159.5

20 mM Tris/HCl, 50 mM ammonium sulfate, 0.1 MM MnCl2

45 °C

1 mM L-Malic acid

Carbon Dioxide (CO2), Pyruvate

0.05 mM NADP+

—

Classical aqueous control (in %)

Activity - Classical

Activity measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm) in the presence of organic solvent

Methanol

30%

100.0

136.8

20 mM Tris/HCl, 50 mM ammonium sulfate, 0.1 MM MnCl2

45 °C

1 mM L-Malic acid

Carbon Dioxide (CO2), Pyruvate

0.05 mM NADP+

—

Classical aqueous control (in %)

Activity - Classical

Activity measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm) in the presence of organic solvent

Methanol

10%

100.0

118.2

20 mM Tris/HCl, 50 mM ammonium sulfate, 0.1 MM MnCl2

45 °C

1 mM L-Malic acid

Carbon Dioxide (CO2), Pyruvate

0.05 mM NADP+

—

Classical aqueous control (in %)

Activity - Classical

Activity measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm) in the presence of organic solvent

Dimethylformamide (DMF)

50%

100.0

101.4

20 mM Tris/HCl, 50 mM ammonium sulfate, 0.1 MM MnCl2

25 °C

1 mM L-Malic acid

Carbon Dioxide (CO2), Pyruvate

0.05 mM NADP+

—

Classical aqueous control (in %)

Activity - Classical

Activity measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm) in the presence of organic solvent

Dimethylformamide (DMF)

30%

100.0

106.8

20 mM Tris/HCl, 50 mM ammonium sulfate, 0.1 MM MnCl2

25 °C

1 mM L-Malic acid

Carbon Dioxide (CO2), Pyruvate

0.05 mM NADP+

—

Classical aqueous control (in %)

Activity - Classical

Activity measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm) in the presence of organic solvent

Dimethylformamide (DMF)

10%

100.0

111.5

20 mM Tris/HCl, 50 mM ammonium sulfate, 0.1 MM MnCl2

25 °C

1 mM L-Malic acid

Carbon Dioxide (CO2), Pyruvate

0.05 mM NADP+

—

Classical aqueous control (in %)

Activity - Classical

Activity measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm) in the presence of organic solvent

Isopropanol

50%

100.0

109.3

20 mM Tris/HCl, 50 mM ammonium sulfate, 0.1 MM MnCl2

25 °C

1 mM L-Malic acid

Carbon Dioxide (CO2), Pyruvate

0.05 mM NADP+

—

Classical aqueous control (in %)

Activity - Classical

Activity measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm) in the presence of organic solvent

Isopropanol

30%

100.0

110.7

20 mM Tris/HCl, 50 mM ammonium sulfate, 0.1 MM MnCl2

25 °C

1 mM L-Malic acid

Carbon Dioxide (CO2), Pyruvate

0.05 mM NADP+

—

Classical aqueous control (in %)

Activity - Classical

Activity measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm) in the presence of organic solvent

Isopropanol

20%

100.0

112.6

20 mM Tris/HCl, 50 mM ammonium sulfate, 0.1 MM MnCl2

25 °C

1 mM L-Malic acid

Carbon Dioxide (CO2), Pyruvate

0.05 mM NADP+

—

Classical aqueous control (in %)

Activity - Classical

Activity measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm) in the presence of organic solvent

Isopropanol

10%

100.0

114.2

20 mM Tris/HCl, 50 mM ammonium sulfate, 0.1 MM MnCl2

25 °C

1 mM L-Malic acid

Carbon Dioxide (CO2), Pyruvate

0.05 mM NADP+

—

Classical aqueous control (in %)

Activity - Classical

Activity measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm) in the presence of organic solvent

Acetone

40%

100.0

139.4

20 mM Tris/HCl, 50 mM ammonium sulfate, 0.1 MM MnCl2

25 °C

1 mM L-Malic acid

Carbon Dioxide (CO2), Pyruvate

0.05 mM NADP+

—

Classical aqueous control (in %)

Stability - Incubation

Activity after incubation (1 hour at 25°C), measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm) in aqueous phase

Tetrahydrofuran (THF)

50%

100.0

50.7

Incubation: 20 mM Tris/HCl buffer, Assay: 50 mM ammonium sulfate, 0.1 MM MnCl2

Incubation: 8, Assay: 8

Incubation: 25°C, Assay: 60°C

1 mM L-Malic acid

Carbon Dioxide (CO2), Pyruvate

Assay: 0.05 mM NADP+

—

Non-incubated control (in %), assay in aqueous phase | Clear about assay conditions

Stability - Incubation

Activity after incubation (24 hours at 25°C), measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm) in aqueous phase

Tetrahydrofuran (THF)

50%

100.0

0.0

Incubation: 20 mM Tris/HCl buffer, Assay: 50 mM ammonium sulfate, 0.1 MM MnCl2

Incubation: 8, Assay: 8

Incubation: 25°C, Assay: 60°C

1 mM L-Malic acid

Carbon Dioxide (CO2), Pyruvate

Assay: 0.05 mM NADP+

—

Non-incubated control (in %), assay in aqueous phase | Clear about assay conditions

Stability - Incubation

Activity after incubation (20 hours at 25°C), measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm) in aqueous phase

Tetrahydrofuran (THF)

50%

100.0

0.0

Incubation: 20 mM Tris/HCl buffer, Assay: 50 mM ammonium sulfate, 0.1 MM MnCl2

Incubation: 8, Assay: 8

Incubation: 25°C, Assay: 60°C

1 mM L-Malic acid

Carbon Dioxide (CO2), Pyruvate

Assay: 0.05 mM NADP+

—

Non-incubated control (in %), assay in aqueous phase | Clear about assay conditions

Stability - Incubation

Activity after incubation (8 hours at 25°C), measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm) in aqueous phase

Tetrahydrofuran (THF)

50%

100.0

16.4

Incubation: 20 mM Tris/HCl buffer, Assay: 50 mM ammonium sulfate, 0.1 MM MnCl2

Incubation: 8, Assay: 8

Incubation: 25°C, Assay: 60°C

1 mM L-Malic acid

Carbon Dioxide (CO2), Pyruvate

Assay: 0.05 mM NADP+

—

Non-incubated control (in %), assay in aqueous phase | Clear about assay conditions

Stability - Incubation

Activity after incubation (4 hours at 25°C), measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm) in aqueous phase

Tetrahydrofuran (THF)

50%

100.0

26.8

Incubation: 20 mM Tris/HCl buffer, Assay: 50 mM ammonium sulfate, 0.1 MM MnCl2

Incubation: 8, Assay: 8

Incubation: 25°C, Assay: 60°C

1 mM L-Malic acid

Carbon Dioxide (CO2), Pyruvate

Assay: 0.05 mM NADP+

—

Non-incubated control (in %), assay in aqueous phase | Clear about assay conditions

Stability - Incubation

Activity after incubation (2 hours at 25°C), measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm) in aqueous phase

Tetrahydrofuran (THF)

50%

100.0

33.9

Incubation: 20 mM Tris/HCl buffer, Assay: 50 mM ammonium sulfate, 0.1 MM MnCl2

Incubation: 8, Assay: 8

Incubation: 25°C, Assay: 60°C

1 mM L-Malic acid

Carbon Dioxide (CO2), Pyruvate

Assay: 0.05 mM NADP+

—

Non-incubated control (in %), assay in aqueous phase | Clear about assay conditions

Stability - Incubation

Activity after incubation (20 hours at 25°C), measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm) in aqueous phase

Dimethylformamide (DMF)

50%

100.0

98.2

Incubation: 20 mM Tris/HCl buffer, Assay: 50 mM ammonium sulfate, 0.1 MM MnCl2

Incubation: 8, Assay: 8

Incubation: 25°C, Assay: 60°C

1 mM L-Malic acid

Carbon Dioxide (CO2), Pyruvate

Assay: 0.05 mM NADP+

—

Non-incubated control (in %), assay in aqueous phase | Clear about assay conditions

Stability - Incubation

Activity after incubation (1 hour at 25°C), measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm) in aqueous phase

Ethanol

15%

100.0

95.0

Incubation: 20 mM Tris/HCl buffer, Assay: 50 mM ammonium sulfate, 0.1 MM MnCl2

Incubation: 8, Assay: 8

Incubation: 25°C, Assay: 60°C

1 mM L-Malic acid

Carbon Dioxide (CO2), Pyruvate

Assay: 0.05 mM NADP+

—

Non-incubated control (in %), assay in aqueous phase | Clear about assay conditions

Stability - Incubation

Activity after incubation (24 hours at 25°C), measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm) in aqueous phase

Methanol

50%

100.0

84.3

Incubation: 20 mM Tris/HCl buffer, Assay: 50 mM ammonium sulfate, 0.1 MM MnCl2

Incubation: 8, Assay: 8

Incubation: 25°C, Assay: 60°C

1 mM L-Malic acid

Carbon Dioxide (CO2), Pyruvate

Assay: 0.05 mM NADP+

—

Non-incubated control (in %), assay in aqueous phase | Clear about assay conditions

Stability - Incubation

Activity after incubation (20 hours at 25°C), measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm) in aqueous phase

Methanol

50%

100.0

85.7

Incubation: 20 mM Tris/HCl buffer, Assay: 50 mM ammonium sulfate, 0.1 MM MnCl2

Incubation: 8, Assay: 8

Incubation: 25°C, Assay: 60°C

1 mM L-Malic acid

Carbon Dioxide (CO2), Pyruvate

Assay: 0.05 mM NADP+

—

Non-incubated control (in %), assay in aqueous phase | Clear about assay conditions

Stability - Incubation

Activity after incubation (6 hours at 25°C), measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm) in aqueous phase

Methanol

50%

100.0

85.7

Incubation: 20 mM Tris/HCl buffer, Assay: 50 mM ammonium sulfate, 0.1 MM MnCl2

Incubation: 8, Assay: 8

Incubation: 25°C, Assay: 60°C

1 mM L-Malic acid

Carbon Dioxide (CO2), Pyruvate

Assay: 0.05 mM NADP+

—

Non-incubated control (in %), assay in aqueous phase | Clear about assay conditions

Stability - Incubation

Activity after incubation (4 hours at 25°C), measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm) in aqueous phase

Methanol

50%

100.0

85.4

Incubation: 20 mM Tris/HCl buffer, Assay: 50 mM ammonium sulfate, 0.1 MM MnCl2

Incubation: 8, Assay: 8

Incubation: 25°C, Assay: 60°C

1 mM L-Malic acid

Carbon Dioxide (CO2), Pyruvate

Assay: 0.05 mM NADP+

—

Non-incubated control (in %), assay in aqueous phase | Clear about assay conditions

Stability - Incubation

Activity after incubation (2 hours at 25°C ), measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm) in aqueous phase

Methanol

50%

100.0

85.0

Incubation: 20 mM Tris/HCl buffer, Assay: 50 mM ammonium sulfate, 0.1 MM MnCl2

Incubation: 8, Assay: 8

Incubation: 25°C, Assay: 60°C

1 mM L-Malic acid

Carbon Dioxide (CO2), Pyruvate

Assay: 0.05 mM NADP+

—

Non-incubated control (in %), assay in aqueous phase | Clear about assay conditions

Stability - Incubation

Activity after incubation (1 hour at 25°C), measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm) in aqueous phase

Methanol

50%

100.0

90.0

Incubation: 20 mM Tris/HCl buffer, Assay: 50 mM ammonium sulfate, 0.1 MM MnCl2

Incubation: 8, Assay: 8

Incubation: 25°C, Assay: 60°C

1 mM L-Malic acid

Carbon Dioxide (CO2), Pyruvate

Assay: 0.05 mM NADP+

—

Non-incubated control (in %), assay in aqueous phase | Clear about assay conditions

Stability - Incubation

Activity after incubation (24 hours at 25°C), measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm) in aqueous phase

Dimethylformamide (DMF)

50%

100.0

97.8

Incubation: 20 mM Tris/HCl buffer, Assay: 50 mM ammonium sulfate, 0.1 MM MnCl2

Incubation: 8, Assay: 8

Incubation: 25°C, Assay: 60°C

1 mM L-Malic acid

Carbon Dioxide (CO2), Pyruvate

Assay: 0.05 mM NADP+

—

Non-incubated control (in %), assay in aqueous phase | Clear about assay conditions

Stability - Incubation

Activity after incubation (2 hours at 25°C), measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm) in aqueous phase

Ethanol

15%

100.0

89.3

Incubation: 20 mM Tris/HCl buffer, Assay: 50 mM ammonium sulfate, 0.1 MM MnCl2

Incubation: 8, Assay: 8

Incubation: 25°C, Assay: 60°C

1 mM L-Malic acid

Carbon Dioxide (CO2), Pyruvate

Assay: 0.05 mM NADP+

—

Non-incubated control (in %), assay in aqueous phase | Clear about assay conditions

Stability - Incubation

Activity after incubation (8 hours at 25°C), measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm) in aqueous phase

Dimethylformamide (DMF)

50%

100.0

101.8

Incubation: 20 mM Tris/HCl buffer, Assay: 50 mM ammonium sulfate, 0.1 MM MnCl2

Incubation: 8, Assay: 8

Incubation: 25°C, Assay: 60°C

1 mM L-Malic acid

Carbon Dioxide (CO2), Pyruvate

Assay: 0.05 mM NADP+

—

Non-incubated control (in %), assay in aqueous phase | Clear about assay conditions

Stability - Incubation

Activity after incubation (4 hours at 25°C), measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm) in aqueous phase

Dimethylformamide (DMF)

50%

100.0

102.1

Incubation: 20 mM Tris/HCl buffer, Assay: 50 mM ammonium sulfate, 0.1 MM MnCl2

Incubation: 8, Assay: 8

Incubation: 25°C, Assay: 60°C

1 mM L-Malic acid

Carbon Dioxide (CO2), Pyruvate

Assay: 0.05 mM NADP+

—

Non-incubated control (in %), assay in aqueous phase | Clear about assay conditions

Stability - Incubation

Activity after incubation (2 hours at 25°C), measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm) in aqueous phase

Dimethylformamide (DMF)

50%

100.0

102.8

Incubation: 20 mM Tris/HCl buffer, Assay: 50 mM ammonium sulfate, 0.1 MM MnCl2

Incubation: 8, Assay: 8

Incubation: 25°C, Assay: 60°C

1 mM L-Malic acid

Carbon Dioxide (CO2), Pyruvate

Assay: 0.05 mM NADP+

—

Non-incubated control (in %), assay in aqueous phase | Clear about assay conditions

Stability - Incubation

Activity after incubation (24 hours at 25°C), measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm) in aqueous phase

Isopropanol

10%

100.0

115.7

Incubation: 20 mM Tris/HCl buffer, Assay: 50 mM ammonium sulfate, 0.1 MM MnCl2

Incubation: 8, Assay: 8

Incubation: 25°C, Assay: 60°C

1 mM L-Malic acid

Carbon Dioxide (CO2), Pyruvate

Assay: 0.05 mM NADP+

—

Non-incubated control (in %), assay in aqueous phase | Clear about assay conditions

Stability - Incubation

Activity after incubation (20 hours at 25°C), measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm) in aqueous phase

Isopropanol

10%

100.0

115.7

Incubation: 20 mM Tris/HCl buffer, Assay: 50 mM ammonium sulfate, 0.1 MM MnCl2

Incubation: 8, Assay: 8

Incubation: 25°C, Assay: 60°C

1 mM L-Malic acid

Carbon Dioxide (CO2), Pyruvate

Assay: 0.05 mM NADP+

—

Non-incubated control (in %), assay in aqueous phase | Clear about assay conditions

Stability - Incubation

Activity after incubation (6 hours at 25°C), measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm) in aqueous phase

Isopropanol

10%

100.0

119.3

Incubation: 20 mM Tris/HCl buffer, Assay: 50 mM ammonium sulfate, 0.1 MM MnCl2

Incubation: 8, Assay: 8

Incubation: 25°C, Assay: 60°C

1 mM L-Malic acid

Carbon Dioxide (CO2), Pyruvate

Assay: 0.05 mM NADP+

—

Non-incubated control (in %), assay in aqueous phase | Clear about assay conditions

Stability - Incubation

Activity after incubation (4 hours at 25°C), measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm) in aqueous phase

Isopropanol

10%

100.0

113.6

Incubation: 20 mM Tris/HCl buffer, Assay: 50 mM ammonium sulfate, 0.1 MM MnCl2

Incubation: 8, Assay: 8

Incubation: 25°C, Assay: 60°C

1 mM L-Malic acid

Carbon Dioxide (CO2), Pyruvate

Assay: 0.05 mM NADP+

—

Non-incubated control (in %), assay in aqueous phase | Clear about assay conditions

Stability - Incubation

Activity after incubation (2 hours at 25°C), measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm) in aqueous phase

Isopropanol

10%

100.0

109.6

Incubation: 20 mM Tris/HCl buffer, Assay: 50 mM ammonium sulfate, 0.1 MM MnCl2

Incubation: 8, Assay: 8

Incubation: 25°C, Assay: 60°C

1 mM L-Malic acid

Carbon Dioxide (CO2), Pyruvate

Assay: 0.05 mM NADP+

—

Non-incubated control (in %), assay in aqueous phase | Clear about assay conditions

Stability - Incubation

Activity after incubation (20 hours at 25°C), measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm) in aqueous phase

Isopropanol

30%

100.0

29.3

Incubation: 20 mM Tris/HCl buffer, Assay: 50 mM ammonium sulfate, 0.1 MM MnCl2

Incubation: 8, Assay: 8

Incubation: 25°C, Assay: 60°C

1 mM L-Malic acid

Carbon Dioxide (CO2), Pyruvate

Assay: 0.05 mM NADP+

—

Non-incubated control (in %), assay in aqueous phase | Clear about assay conditions

Stability - Incubation

Activity after incubation (0.5 hour at 25°C), measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm) in aqueous phase

Tetrahydrofuran (THF)

50%

100.0

82.1

Incubation: 20 mM Tris/HCl buffer, Assay: 50 mM ammonium sulfate, 0.1 MM MnCl2

Incubation: 8, Assay: 8

Incubation: 25°C, Assay: 60°C

1 mM L-Malic acid

Carbon Dioxide (CO2), Pyruvate

Assay: 0.05 mM NADP+

—

Non-incubated control (in %), assay in aqueous phase | Clear about assay conditions

Stability - Incubation

Activity after incubation (24 hours at 25°C), measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm) in aqueous phase

Ethanol

50%

100.0

15.7

Incubation: 20 mM Tris/HCl buffer, Assay: 50 mM ammonium sulfate, 0.1 MM MnCl2

Incubation: 8, Assay: 8

Incubation: 25°C, Assay: 60°C

1 mM L-Malic acid

Carbon Dioxide (CO2), Pyruvate

Assay: 0.05 mM NADP+

—

Non-incubated control (in %), assay in aqueous phase | Clear about assay conditions

Stability - Incubation

Activity after incubation (20 hours at 25°C), measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm) in aqueous phase

Ethanol

50%

100.0

22.1

Incubation: 20 mM Tris/HCl buffer, Assay: 50 mM ammonium sulfate, 0.1 MM MnCl2

Incubation: 8, Assay: 8

Incubation: 25°C, Assay: 60°C

1 mM L-Malic acid

Carbon Dioxide (CO2), Pyruvate

Assay: 0.05 mM NADP+

—

Non-incubated control (in %), assay in aqueous phase | Clear about assay conditions

Stability - Incubation

Activity after incubation (8 hours at 25°C), measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm) in aqueous phase

Ethanol

50%

100.0

50.0

Incubation: 20 mM Tris/HCl buffer, Assay: 50 mM ammonium sulfate, 0.1 MM MnCl2

Incubation: 8, Assay: 8

Incubation: 25°C, Assay: 60°C

1 mM L-Malic acid

Carbon Dioxide (CO2), Pyruvate

Assay: 0.05 mM NADP+

—

Non-incubated control (in %), assay in aqueous phase | Clear about assay conditions

Stability - Incubation

Activity after incubation (4 hours at 25°C), measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm) in aqueous phase

Ethanol

50%

100.0

57.1

Incubation: 20 mM Tris/HCl buffer, Assay: 50 mM ammonium sulfate, 0.1 MM MnCl2

Incubation: 8, Assay: 8

Incubation: 25°C, Assay: 60°C

1 mM L-Malic acid

Carbon Dioxide (CO2), Pyruvate

Assay: 0.05 mM NADP+

—

Non-incubated control (in %), assay in aqueous phase | Clear about assay conditions

Stability - Incubation

Activity after incubation (2 hours at 25°C), measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm) in aqueous phase

Ethanol

50%

100.0

64.3

Incubation: 20 mM Tris/HCl buffer, Assay: 50 mM ammonium sulfate, 0.1 MM MnCl2

Incubation: 8, Assay: 8

Incubation: 25°C, Assay: 60°C

1 mM L-Malic acid

Carbon Dioxide (CO2), Pyruvate

Assay: 0.05 mM NADP+

—

Non-incubated control (in %), assay in aqueous phase | Clear about assay conditions

Stability - Incubation

Activity after incubation (1 hour at 25°C), measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm) in aqueous phase

Ethanol

50%

100.0

76.4

Incubation: 20 mM Tris/HCl buffer, Assay: 50 mM ammonium sulfate, 0.1 MM MnCl2

Incubation: 8, Assay: 8

Incubation: 25°C, Assay: 60°C

1 mM L-Malic acid

Carbon Dioxide (CO2), Pyruvate

Assay: 0.05 mM NADP+

—

Non-incubated control (in %), assay in aqueous phase | Clear about assay conditions

Stability - Incubation

Activity after incubation (24 hours at 25°C), measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm) in aqueous phase

Isopropanol

30%

100.0

20.7

Incubation: 20 mM Tris/HCl buffer, Assay: 50 mM ammonium sulfate, 0.1 MM MnCl2

Incubation: 8, Assay: 8

Incubation: 25°C, Assay: 60°C

1 mM L-Malic acid

Carbon Dioxide (CO2), Pyruvate

Assay: 0.05 mM NADP+

—

Non-incubated control (in %), assay in aqueous phase | Clear about assay conditions

Stability - Incubation

Activity after incubation (1 hour at 25°C), measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm) in aqueous phase

Isopropanol

10%

100.0

108.2

Incubation: 20 mM Tris/HCl buffer, Assay: 50 mM ammonium sulfate, 0.1 MM MnCl2

Incubation: 8, Assay: 8

Incubation: 25°C, Assay: 60°C

1 mM L-Malic acid

Carbon Dioxide (CO2), Pyruvate

Assay: 0.05 mM NADP+

—

Non-incubated control (in %), assay in aqueous phase | Clear about assay conditions

Stability - Incubation

Activity after incubation (8 hours at 25°C), measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm) in aqueous phase

Isopropanol

30%

100.0

58.9

Incubation: 20 mM Tris/HCl buffer, Assay: 50 mM ammonium sulfate, 0.1 MM MnCl2

Incubation: 8, Assay: 8

Incubation: 25°C, Assay: 60°C

1 mM L-Malic acid

Carbon Dioxide (CO2), Pyruvate

Assay: 0.05 mM NADP+

—

Non-incubated control (in %), assay in aqueous phase | Clear about assay conditions

Stability - Incubation

Activity after incubation (4 hours at 25°C), measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm) in aqueous phase

Isopropanol

30%

100.0

66.4

Incubation: 20 mM Tris/HCl buffer, Assay: 50 mM ammonium sulfate, 0.1 MM MnCl2

Incubation: 8, Assay: 8

Incubation: 25°C, Assay: 60°C

1 mM L-Malic acid

Carbon Dioxide (CO2), Pyruvate

Assay: 0.05 mM NADP+

—

Non-incubated control (in %), assay in aqueous phase | Clear about assay conditions

Stability - Incubation

Activity after incubation (2 hours at 25°C), measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm) in aqueous phase

Isopropanol

30%

100.0

80.7

Incubation: 20 mM Tris/HCl buffer, Assay: 50 mM ammonium sulfate, 0.1 MM MnCl2

Incubation: 8, Assay: 8

Incubation: 25°C, Assay: 60°C

1 mM L-Malic acid

Carbon Dioxide (CO2), Pyruvate

Assay: 0.05 mM NADP+

—

Non-incubated control (in %), assay in aqueous phase | Clear about assay conditions

Stability - Incubation

Activity after incubation (24 hours at 25°C), measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm) in aqueous phase

Ethanol

15%

100.0

85.4

Incubation: 20 mM Tris/HCl buffer, Assay: 50 mM ammonium sulfate, 0.1 MM MnCl2

Incubation: 8, Assay: 8

Incubation: 25°C, Assay: 60°C

1 mM L-Malic acid

Carbon Dioxide (CO2), Pyruvate

Assay: 0.05 mM NADP+

—

Non-incubated control (in %), assay in aqueous phase | Clear about assay conditions

Stability - Incubation

Activity after incubation (20 hours at 25°C), measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm) in aqueous phase

Ethanol

15%

100.0

85.7

Incubation: 20 mM Tris/HCl buffer, Assay: 50 mM ammonium sulfate, 0.1 MM MnCl2

Incubation: 8, Assay: 8

Incubation: 25°C, Assay: 60°C

1 mM L-Malic acid

Carbon Dioxide (CO2), Pyruvate

Assay: 0.05 mM NADP+

—

Non-incubated control (in %), assay in aqueous phase | Clear about assay conditions

Stability - Incubation

Activity after incubation (6 hours at 25°C), measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm) in aqueous phase

Ethanol

15%

100.0

86.1

Incubation: 20 mM Tris/HCl buffer, Assay: 50 mM ammonium sulfate, 0.1 MM MnCl2

Incubation: 8, Assay: 8

Incubation: 25°C, Assay: 60°C

1 mM L-Malic acid

Carbon Dioxide (CO2), Pyruvate

Assay: 0.05 mM NADP+

—

Non-incubated control (in %), assay in aqueous phase | Clear about assay conditions

Stability - Incubation

Activity after incubation (4 hours at 25°C), measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm) in aqueous phase

Ethanol

15%

100.0

87.1

Incubation: 20 mM Tris/HCl buffer, Assay: 50 mM ammonium sulfate, 0.1 MM MnCl2

Incubation: 8, Assay: 8

Incubation: 25°C, Assay: 60°C

1 mM L-Malic acid

Carbon Dioxide (CO2), Pyruvate

Assay: 0.05 mM NADP+

—

Non-incubated control (in %), assay in aqueous phase | Clear about assay conditions

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Visualization : Activity — Classical

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

Visualization : Stability — Incubation

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

Structure

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NADP-dependent malic enzyme from Sulfolobus solfataricus

Enzyme Description

Sequence

Experimental Data (71 measurements)

Visualization : Activity — Classical

Visualization : Stability — Incubation

Structure

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