Ene-reductase DrER from Deinococcus radiodurans

Enzyme Description

Species

Deinococcus radiodurans ↗

Extremophile

Yes polyextremophilic organism

EC Number

1.6.99.1 ↗

Sequence

Length: 370 amino acids

MTVSSAAAPQPASPAAPLLFTPLKLRSLELPNRVVVSPMCTYSATDGVANEFHLVHLGQYALGGAGLILAEATAVSPEGRITPEDLGLWDDRQIVPLGHITDFVHQHGGHIGVQLAHAGRKASTYAPWRGKGAVPAELGGWQVIGPDENSFHDLFPTPAMMGADELRGVVDAFSAAARRAQVAGFDAVEVHAAHGYLLHQFLSPLANTRTDDYGGSFENRTRLLLEVVRAVRHVWPAHLPLFVRLSATDWAEGGWDLEQTVQLSKLLKYEGVDVLDISSGGLTAAQQIEVGPGYQVPFAAAVSRAETEISVMAVGLIETGAQAEAILQAGDADLIALGRPFLRDPHWAQRAARELGLRPVSIDQYARAGW

Frieda A. Sorgenfrei et al. (2024) — Solvent concentration at 50% protein unfolding may reform enzyme stability ranking and process window identification

Nature Communications · doi:10.1038/s41467-024-49774-0 ↗ · Activity - Classical Stability - C50 Stability - Tm

164 measurements ▼

Database ID

UniProt: Q9RSD4 ↗

Sequence Annotation

Explicit - Provided UniProt Accession Number

Protein Source

Recombinant, host bacterium Escherichia coli BL21 (DE3)

Experimental Data (164 measurements)

164 measurements — page 1 of 9

Property	Assay	Solvent	Solvent Volume	Aqueous Reference	Measured Value	Units	Extraction method	Data type	Solution	pH	Temperature	Substrate(s)	Product(s)	Cofactor(s)	Shaking	Comments
Activity - Classical	Activity measured by absorbance spectrophotometry (NAD(P)H absorbance measurement, 340 nm) in the presence of organic solvent	Dimethyl Sulfoxide (DMSO)	5 % v/v	100.0	105.4	%	Direct	Continuous	50 mM sodium phosphate	7.4	Room temperature	10 mM Cyclohex-2-enone	Cyclohexane	0.2 mM NAD(P)H	—	Classical aqueous control (in %) \| Raw activity data
Activity - Classical	Activity measured by absorbance spectrophotometry (NAD(P)H absorbance measurement, 340 nm) in the presence of organic solvent	Dimethyl Sulfoxide (DMSO)	10 % v/v	100.0	104.0	%	Direct	Continuous	50 mM sodium phosphate	7.4	Room temperature	10 mM Cyclohex-2-enone	Cyclohexane	0.2 mM NAD(P)H	—	Classical aqueous control (in %) \| Raw activity data
Activity - Classical	Activity measured by absorbance spectrophotometry (NAD(P)H absorbance measurement, 340 nm) in the presence of organic solvent	Dimethyl Sulfoxide (DMSO)	15 % v/v	100.0	99.3	%	Direct	Continuous	50 mM sodium phosphate	7.4	Room temperature	10 mM Cyclohex-2-enone	Cyclohexane	0.2 mM NAD(P)H	—	Classical aqueous control (in %) \| Raw activity data
Activity - Classical	Activity measured by absorbance spectrophotometry (NAD(P)H absorbance measurement, 340 nm) in the presence of organic solvent	Dimethyl Sulfoxide (DMSO)	20 % v/v	100.0	81.9	%	Direct	Continuous	50 mM sodium phosphate	7.4	Room temperature	10 mM Cyclohex-2-enone	Cyclohexane	0.2 mM NAD(P)H	—	Classical aqueous control (in %) \| Raw activity data
Activity - Classical	Activity measured by absorbance spectrophotometry (NAD(P)H absorbance measurement, 340 nm) in the presence of organic solvent	Dimethyl Sulfoxide (DMSO)	25 % v/v	100.0	69.1	%	Direct	Continuous	50 mM sodium phosphate	7.4	Room temperature	10 mM Cyclohex-2-enone	Cyclohexane	0.2 mM NAD(P)H	—	Classical aqueous control (in %) \| Raw activity data
Activity - Classical	Activity measured by absorbance spectrophotometry (NAD(P)H absorbance measurement, 340 nm) in the presence of organic solvent	Dimethyl Sulfoxide (DMSO)	30 % v/v	100.0	47.7	%	Direct	Continuous	50 mM sodium phosphate	7.4	Room temperature	10 mM Cyclohex-2-enone	Cyclohexane	0.2 mM NAD(P)H	—	Classical aqueous control (in %) \| Raw activity data
Activity - Classical	Activity measured by absorbance spectrophotometry (NAD(P)H absorbance measurement, 340 nm) in the presence of organic solvent	Ethanol	5 % v/v	100.0	84.0	%	Direct	Continuous	50 mM sodium phosphate	7.4	25°C	10 mM Cyclohex-2-enone	Cyclohexane	0.2 mM NAD(P)H	—	Classical aqueous control (in %) \| Raw activity data
Activity - Classical	Activity measured by absorbance spectrophotometry (NAD(P)H absorbance measurement, 340 nm) in the presence of organic solvent	Ethanol	10 % v/v	100.0	78.8	%	Direct	Continuous	50 mM sodium phosphate	7.4	25°C	10 mM Cyclohex-2-enone	Cyclohexane	0.2 mM NAD(P)H	—	Classical aqueous control (in %) \| Raw activity data
Activity - Classical	Activity measured by absorbance spectrophotometry (NAD(P)H absorbance measurement, 340 nm) in the presence of organic solvent	Ethanol	15 % v/v	100.0	89.7	%	Direct	Continuous	50 mM sodium phosphate	7.4	25°C	10 mM Cyclohex-2-enone	Cyclohexane	0.2 mM NAD(P)H	—	Classical aqueous control (in %) \| Raw activity data
Activity - Classical	Activity measured by absorbance spectrophotometry (NAD(P)H absorbance measurement, 340 nm) in the presence of organic solvent	Ethanol	20 % v/v	100.0	82.1	%	Direct	Continuous	50 mM sodium phosphate	7.4	25°C	10 mM Cyclohex-2-enone	Cyclohexane	0.2 mM NAD(P)H	—	Classical aqueous control (in %) \| Raw activity data
Activity - Classical	Activity measured by absorbance spectrophotometry (NAD(P)H absorbance measurement, 340 nm) in the presence of organic solvent	Ethanol	25 % v/v	100.0	53.8	%	Direct	Continuous	50 mM sodium phosphate	7.4	25°C	10 mM Cyclohex-2-enone	Cyclohexane	0.2 mM NAD(P)H	—	Classical aqueous control (in %) \| Raw activity data
Activity - Classical	Activity measured by absorbance spectrophotometry (NAD(P)H absorbance measurement, 340 nm) in the presence of organic solvent	Ethanol	30 % v/v	100.0	26.9	%	Direct	Continuous	50 mM sodium phosphate	7.4	25°C	10 mM Cyclohex-2-enone	Cyclohexane	0.2 mM NAD(P)H	—	Classical aqueous control (in %) \| Raw activity data
Activity - Classical	Activity measured by absorbance spectrophotometry (NAD(P)H absorbance measurement, 340 nm) in the presence of organic solvent	Methanol	5 % v/v	100.0	91.5	%	Direct	Continuous	50 mM sodium phosphate	7.4	25°C	10 mM Cyclohex-2-enone	Cyclohexane	0.2 mM NAD(P)H	—	Classical aqueous control (in %) \| Raw activity data
Activity - Classical	Activity measured by absorbance spectrophotometry (NAD(P)H absorbance measurement, 340 nm) in the presence of organic solvent	Methanol	10 % v/v	100.0	85.9	%	Direct	Continuous	50 mM sodium phosphate	7.4	25°C	10 mM Cyclohex-2-enone	Cyclohexane	0.2 mM NAD(P)H	—	Classical aqueous control (in %) \| Raw activity data
Activity - Classical	Activity measured by absorbance spectrophotometry (NAD(P)H absorbance measurement, 340 nm) in the presence of organic solvent	Methanol	15 % v/v	100.0	81.7	%	Direct	Continuous	50 mM sodium phosphate	7.4	25°C	10 mM Cyclohex-2-enone	Cyclohexane	0.2 mM NAD(P)H	—	Classical aqueous control (in %) \| Raw activity data
Activity - Classical	Activity measured by absorbance spectrophotometry (NAD(P)H absorbance measurement, 340 nm) in the presence of organic solvent	Methanol	20 % v/v	100.0	77.5	%	Direct	Continuous	50 mM sodium phosphate	7.4	25°C	10 mM Cyclohex-2-enone	Cyclohexane	0.2 mM NAD(P)H	—	Classical aqueous control (in %) \| Raw activity data
Activity - Classical	Activity measured by absorbance spectrophotometry (NAD(P)H absorbance measurement, 340 nm) in the presence of organic solvent	Methanol	25 % v/v	100.0	57.0	%	Direct	Continuous	50 mM sodium phosphate	7.4	25°C	10 mM Cyclohex-2-enone	Cyclohexane	0.2 mM NAD(P)H	—	Classical aqueous control (in %) \| Raw activity data
Activity - Classical	Activity measured by absorbance spectrophotometry (NAD(P)H absorbance measurement, 340 nm) in the presence of organic solvent	Methanol	30 % v/v	100.0	27.5	%	Direct	Continuous	50 mM sodium phosphate	7.4	25°C	10 mM Cyclohex-2-enone	Cyclohexane	0.2 mM NAD(P)H	—	Classical aqueous control (in %) \| Raw activity data
Activity - Classical	Activity measured by absorbance spectrophotometry (NAD(P)H absorbance measurement, 340 nm) in the presence of organic solvent	Isopropanol	5 % v/v	100.0	94.2	%	Direct	Continuous	50 mM sodium phosphate	7.4	Room temperature	10 mM Cyclohex-2-enone	Cyclohexane	0.2 mM NAD(P)H	—	Classical aqueous control (in %) \| Raw activity data
Activity - Classical	Activity measured by absorbance spectrophotometry (NAD(P)H absorbance measurement, 340 nm) in the presence of organic solvent	Isopropanol	10 % v/v	100.0	98.7	%	Direct	Continuous	50 mM sodium phosphate	7.4	Room temperature	10 mM Cyclohex-2-enone	Cyclohexane	0.2 mM NAD(P)H	—	Classical aqueous control (in %) \| Raw activity data

Activity - Classical

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

Dimethyl Sulfoxide (DMSO)

5 % v/v

100.0

105.4

Direct

Continuous

50 mM sodium phosphate

7.4

Room temperature

10 mM Cyclohex-2-enone

Cyclohexane

0.2 mM NAD(P)H

—

Classical aqueous control (in %) | Raw activity data

Activity - Classical

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

Dimethyl Sulfoxide (DMSO)

10 % v/v

100.0

104.0

Direct

Continuous

50 mM sodium phosphate

7.4

Room temperature

10 mM Cyclohex-2-enone

Cyclohexane

0.2 mM NAD(P)H

—

Classical aqueous control (in %) | Raw activity data

Activity - Classical

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

Dimethyl Sulfoxide (DMSO)

15 % v/v

100.0

99.3

Direct

Continuous

50 mM sodium phosphate

7.4

Room temperature

10 mM Cyclohex-2-enone

Cyclohexane

0.2 mM NAD(P)H

—

Classical aqueous control (in %) | Raw activity data

Activity - Classical

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

Dimethyl Sulfoxide (DMSO)

20 % v/v

100.0

81.9

Direct

Continuous

50 mM sodium phosphate

7.4

Room temperature

10 mM Cyclohex-2-enone

Cyclohexane

0.2 mM NAD(P)H

—

Classical aqueous control (in %) | Raw activity data

Activity - Classical

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

Dimethyl Sulfoxide (DMSO)

25 % v/v

100.0

69.1

Direct

Continuous

50 mM sodium phosphate

7.4

Room temperature

10 mM Cyclohex-2-enone

Cyclohexane

0.2 mM NAD(P)H

—

Classical aqueous control (in %) | Raw activity data

Activity - Classical

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

Dimethyl Sulfoxide (DMSO)

30 % v/v

100.0

47.7

Direct

Continuous

50 mM sodium phosphate

7.4

Room temperature

10 mM Cyclohex-2-enone

Cyclohexane

0.2 mM NAD(P)H

—

Classical aqueous control (in %) | Raw activity data

Activity - Classical

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

Ethanol

5 % v/v

100.0

84.0

Direct

Continuous

50 mM sodium phosphate

7.4

25°C

10 mM Cyclohex-2-enone

Cyclohexane

0.2 mM NAD(P)H

—

Classical aqueous control (in %) | Raw activity data

Activity - Classical

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

Ethanol

10 % v/v

100.0

78.8

Direct

Continuous

50 mM sodium phosphate

7.4

25°C

10 mM Cyclohex-2-enone

Cyclohexane

0.2 mM NAD(P)H

—

Classical aqueous control (in %) | Raw activity data

Activity - Classical

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

Ethanol

15 % v/v

100.0

89.7

Direct

Continuous

50 mM sodium phosphate

7.4

25°C

10 mM Cyclohex-2-enone

Cyclohexane

0.2 mM NAD(P)H

—

Classical aqueous control (in %) | Raw activity data

Activity - Classical

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

Ethanol

20 % v/v

100.0

82.1

Direct

Continuous

50 mM sodium phosphate

7.4

25°C

10 mM Cyclohex-2-enone

Cyclohexane

0.2 mM NAD(P)H

—

Classical aqueous control (in %) | Raw activity data

Activity - Classical

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

Ethanol

25 % v/v

100.0

53.8

Direct

Continuous

50 mM sodium phosphate

7.4

25°C

10 mM Cyclohex-2-enone

Cyclohexane

0.2 mM NAD(P)H

—

Classical aqueous control (in %) | Raw activity data

Activity - Classical

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

Ethanol

30 % v/v

100.0

26.9

Direct

Continuous

50 mM sodium phosphate

7.4

25°C

10 mM Cyclohex-2-enone

Cyclohexane

0.2 mM NAD(P)H

—

Classical aqueous control (in %) | Raw activity data

Activity - Classical

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

Methanol

5 % v/v

100.0

91.5

Direct

Continuous

50 mM sodium phosphate

7.4

25°C

10 mM Cyclohex-2-enone

Cyclohexane

0.2 mM NAD(P)H

—

Classical aqueous control (in %) | Raw activity data

Activity - Classical

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

Methanol

10 % v/v

100.0

85.9

Direct

Continuous

50 mM sodium phosphate

7.4

25°C

10 mM Cyclohex-2-enone

Cyclohexane

0.2 mM NAD(P)H

—

Classical aqueous control (in %) | Raw activity data

Activity - Classical

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

Methanol

15 % v/v

100.0

81.7

Direct

Continuous

50 mM sodium phosphate

7.4

25°C

10 mM Cyclohex-2-enone

Cyclohexane

0.2 mM NAD(P)H

—

Classical aqueous control (in %) | Raw activity data

Activity - Classical

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

Methanol

20 % v/v

100.0

77.5

Direct

Continuous

50 mM sodium phosphate

7.4

25°C

10 mM Cyclohex-2-enone

Cyclohexane

0.2 mM NAD(P)H

—

Classical aqueous control (in %) | Raw activity data

Activity - Classical

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

Methanol

25 % v/v

100.0

57.0

Direct

Continuous

50 mM sodium phosphate

7.4

25°C

10 mM Cyclohex-2-enone

Cyclohexane

0.2 mM NAD(P)H

—

Classical aqueous control (in %) | Raw activity data

Activity - Classical

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

Methanol

30 % v/v

100.0

27.5

Direct

Continuous

50 mM sodium phosphate

7.4

25°C

10 mM Cyclohex-2-enone

Cyclohexane

0.2 mM NAD(P)H

—

Classical aqueous control (in %) | Raw activity data

Activity - Classical

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

Isopropanol

5 % v/v

100.0

94.2

Direct

Continuous

50 mM sodium phosphate

7.4

Room temperature

10 mM Cyclohex-2-enone

Cyclohexane

0.2 mM NAD(P)H

—

Classical aqueous control (in %) | Raw activity data

Activity - Classical

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

Isopropanol

10 % v/v

100.0

98.7

Direct

Continuous

50 mM sodium phosphate

7.4

Room temperature

10 mM Cyclohex-2-enone

Cyclohexane

0.2 mM NAD(P)H

—

Classical aqueous control (in %) | Raw activity data

Activity - Classical

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

Isopropanol

15 % v/v

100.0

90.4

Direct

Continuous

50 mM sodium phosphate

7.4

Room temperature

10 mM Cyclohex-2-enone

Cyclohexane

0.2 mM NAD(P)H

—

Classical aqueous control (in %) | Raw activity data

Activity - Classical

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

Isopropanol

20 % v/v

100.0

74.4

Direct

Continuous

50 mM sodium phosphate

7.4

Room temperature

10 mM Cyclohex-2-enone

Cyclohexane

0.2 mM NAD(P)H

—

Classical aqueous control (in %) | Raw activity data

Activity - Classical

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

Isopropanol

25 % v/v

100.0

51.9

Direct

Continuous

50 mM sodium phosphate

7.4

Room temperature

10 mM Cyclohex-2-enone

Cyclohexane

0.2 mM NAD(P)H

—

Classical aqueous control (in %) | Raw activity data

Activity - Classical

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

Isopropanol

30 % v/v

100.0

7.7

Direct

Continuous

50 mM sodium phosphate

7.4

Room temperature

10 mM Cyclohex-2-enone

Cyclohexane

0.2 mM NAD(P)H

—

Classical aqueous control (in %) | Raw activity data

Activity - Classical

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

1-Propanol

5 % v/v

100.0

97.8

Direct

Continuous

50 mM sodium phosphate

7.4

Room temperature

10 mM Cyclohex-2-enone

Cyclohexane

0.2 mM NAD(P)H

—

Classical aqueous control (in %) | Raw activity data

Activity - Classical

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

1-Propanol

10 % v/v

100.0

98.5

Direct

Continuous

50 mM sodium phosphate

7.4

Room temperature

10 mM Cyclohex-2-enone

Cyclohexane

0.2 mM NAD(P)H

—

Classical aqueous control (in %) | Raw activity data

Activity - Classical

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

1-Propanol

15 % v/v

100.0

76.9

Direct

Continuous

50 mM sodium phosphate

7.4

Room temperature

10 mM Cyclohex-2-enone

Cyclohexane

0.2 mM NAD(P)H

—

Classical aqueous control (in %) | Raw activity data

Activity - Classical

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

1-Propanol

20 % v/v

100.0

17.9

Direct

Continuous

50 mM sodium phosphate

7.4

Room temperature

10 mM Cyclohex-2-enone

Cyclohexane

0.2 mM NAD(P)H

—

Classical aqueous control (in %) | Raw activity data

Activity - Classical

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

1-Propanol

25 % v/v

100.0

3.0

Direct

Continuous

50 mM sodium phosphate

7.4

Room temperature

10 mM Cyclohex-2-enone

Cyclohexane

0.2 mM NAD(P)H

—

Classical aqueous control (in %) | Raw activity data

Activity - Classical

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

1-Propanol

30 % v/v

100.0

1.5

Direct

Continuous

50 mM sodium phosphate

7.4

Room temperature

10 mM Cyclohex-2-enone

Cyclohexane

0.2 mM NAD(P)H

—

Classical aqueous control (in %) | Raw activity data

Activity - Classical

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

Ethanol

5 % v/v

100.0

93.1

Direct

Continuous

50 mM sodium phosphate

7.4

25°C

10 mM Cyclohex-2-enone

Cyclohexane

0.2 mM NAD(P)H

—

Classical aqueous control (in %) | Raw activity data of reaction temperature screening

Activity - Classical

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

Ethanol

10 % v/v

100.0

145.8

Direct

Continuous

50 mM sodium phosphate

7.4

25°C

10 mM Cyclohex-2-enone

Cyclohexane

0.2 mM NAD(P)H

—

Classical aqueous control (in %) | Raw activity data of reaction temperature screening

Activity - Classical

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

Ethanol

15 % v/v

100.0

136.1

Direct

Continuous

50 mM sodium phosphate

7.4

25°C

10 mM Cyclohex-2-enone

Cyclohexane

0.2 mM NAD(P)H

—

Classical aqueous control (in %) | Raw activity data of reaction temperature screening

Activity - Classical

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

Ethanol

20 % v/v

100.0

94.4

Direct

Continuous

50 mM sodium phosphate

7.4

25°C

10 mM Cyclohex-2-enone

Cyclohexane

0.2 mM NAD(P)H

—

Classical aqueous control (in %) | Raw activity data of reaction temperature screening

Activity - Classical

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

Ethanol

25 % v/v

100.0

51.4

Direct

Continuous

50 mM sodium phosphate

7.4

25°C

10 mM Cyclohex-2-enone

Cyclohexane

0.2 mM NAD(P)H

—

Classical aqueous control (in %) | Raw activity data of reaction temperature screening

Activity - Classical

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

Ethanol

30 % v/v

100.0

25.0

Direct

Continuous

50 mM sodium phosphate

7.4

25°C

10 mM Cyclohex-2-enone

Cyclohexane

0.2 mM NAD(P)H

—

Classical aqueous control (in %) | Raw activity data of reaction temperature screening

Activity - Classical

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

Ethanol

35 % v/v

100.0

19.4

Direct

Continuous

50 mM sodium phosphate

7.4

25°C

10 mM Cyclohex-2-enone

Cyclohexane

0.2 mM NAD(P)H

—

Classical aqueous control (in %) | Raw activity data of reaction temperature screening

Activity - Classical

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

Ethanol

40 % v/v

100.0

5.6

Direct

Continuous

50 mM sodium phosphate

7.4

25°C

10 mM Cyclohex-2-enone

Cyclohexane

0.2 mM NAD(P)H

—

Classical aqueous control (in %) | Raw activity data of reaction temperature screening

Activity - Classical

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

Ethanol

45 % v/v

100.0

15.3

Direct

Continuous

50 mM sodium phosphate

7.4

25°C

10 mM Cyclohex-2-enone

Cyclohexane

0.2 mM NAD(P)H

—

Classical aqueous control (in %) | Raw activity data of reaction temperature screening

Activity - Classical

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

Ethanol

5 % v/v

100.0

103.8

Direct

Continuous

50 mM sodium phosphate

7.4

30°C

10 mM Cyclohex-2-enone

Cyclohexane

0.2 mM NAD(P)H

—

Classical aqueous control (in %) | Raw activity data of reaction temperature screening

Activity - Classical

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

Ethanol

10 % v/v

100.0

107.5

Direct

Continuous

50 mM sodium phosphate

7.4

30°C

10 mM Cyclohex-2-enone

Cyclohexane

0.2 mM NAD(P)H

—

Classical aqueous control (in %) | Raw activity data of reaction temperature screening

Activity - Classical

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

Ethanol

15 % v/v

100.0

94.3

Direct

Continuous

50 mM sodium phosphate

7.4

30°C

10 mM Cyclohex-2-enone

Cyclohexane

0.2 mM NAD(P)H

—

Classical aqueous control (in %) | Raw activity data of reaction temperature screening

Activity - Classical

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

Ethanol

20 % v/v

100.0

41.5

Direct

Continuous

50 mM sodium phosphate

7.4

30°C

10 mM Cyclohex-2-enone

Cyclohexane

0.2 mM NAD(P)H

—

Classical aqueous control (in %) | Raw activity data of reaction temperature screening

Activity - Classical

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

Ethanol

25 % v/v

100.0

9.4

Direct

Continuous

50 mM sodium phosphate

7.4

30°C

10 mM Cyclohex-2-enone

Cyclohexane

0.2 mM NAD(P)H

—

Classical aqueous control (in %) | Raw activity data of reaction temperature screening

Activity - Classical

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

Ethanol

30 % v/v

100.0

5.7

Direct

Continuous

50 mM sodium phosphate

7.4

30°C

10 mM Cyclohex-2-enone

Cyclohexane

0.2 mM NAD(P)H

—

Classical aqueous control (in %) | Raw activity data of reaction temperature screening

Activity - Classical

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

Ethanol

35 % v/v

100.0

0.0

Direct

Continuous

50 mM sodium phosphate

7.4

30°C

10 mM Cyclohex-2-enone

Cyclohexane

0.2 mM NAD(P)H

—

Classical aqueous control (in %) | Raw activity data of reaction temperature screening

Activity - Classical

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

Ethanol

40 % v/v

100.0

1.9

Direct

Continuous

50 mM sodium phosphate

7.4

30°C

10 mM Cyclohex-2-enone

Cyclohexane

0.2 mM NAD(P)H

—

Classical aqueous control (in %) | Raw activity data of reaction temperature screening

Activity - Classical

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

Ethanol

45 % v/v

100.0

9.4

Direct

Continuous

50 mM sodium phosphate

7.4

30°C

10 mM Cyclohex-2-enone

Cyclohexane

0.2 mM NAD(P)H

—

Classical aqueous control (in %) | Raw activity data of reaction temperature screening

Activity - Classical

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

Ethanol

5 % v/v

100.0

78.5

Direct

Continuous

50 mM sodium phosphate

7.4

35°C

10 mM Cyclohex-2-enone

Cyclohexane

0.2 mM NAD(P)H

—

Classical aqueous control (in %) | Raw activity data of reaction temperature screening

Activity - Classical

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

Ethanol

10 % v/v

100.0

85.4

Direct

Continuous

50 mM sodium phosphate

7.4

35°C

10 mM Cyclohex-2-enone

Cyclohexane

0.2 mM NAD(P)H

—

Classical aqueous control (in %) | Raw activity data of reaction temperature screening

Activity - Classical

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

Ethanol

15 % v/v

100.0

39.6

Direct

Continuous

50 mM sodium phosphate

7.4

35°C

10 mM Cyclohex-2-enone

Cyclohexane

0.2 mM NAD(P)H

—

Classical aqueous control (in %) | Raw activity data of reaction temperature screening

Activity - Classical

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

Ethanol

20 % v/v

100.0

10.4

Direct

Continuous

50 mM sodium phosphate

7.4

35°C

10 mM Cyclohex-2-enone

Cyclohexane

0.2 mM NAD(P)H

—

Classical aqueous control (in %) | Raw activity data of reaction temperature screening

Activity - Classical

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

Ethanol

25 % v/v

100.0

1.4

Direct

Continuous

50 mM sodium phosphate

7.4

35°C

10 mM Cyclohex-2-enone

Cyclohexane

0.2 mM NAD(P)H

—

Classical aqueous control (in %) | Raw activity data of reaction temperature screening

Activity - Classical

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

Ethanol

30 % v/v

100.0

3.5

Direct

Continuous

50 mM sodium phosphate

7.4

35°C

10 mM Cyclohex-2-enone

Cyclohexane

0.2 mM NAD(P)H

—

Classical aqueous control (in %) | Raw activity data of reaction temperature screening

Activity - Classical

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

Ethanol

35 % v/v

100.0

2.1

Direct

Continuous

50 mM sodium phosphate

7.4

35°C

10 mM Cyclohex-2-enone

Cyclohexane

0.2 mM NAD(P)H

—

Classical aqueous control (in %) | Raw activity data of reaction temperature screening

Activity - Classical

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

Ethanol

40 % v/v

100.0

2.1

Direct

Continuous

50 mM sodium phosphate

7.4

35°C

10 mM Cyclohex-2-enone

Cyclohexane

0.2 mM NAD(P)H

—

Classical aqueous control (in %) | Raw activity data of reaction temperature screening

Activity - Classical

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

Ethanol

45 % v/v

100.0

0.7

Direct

Continuous

50 mM sodium phosphate

7.4

35°C

10 mM Cyclohex-2-enone

Cyclohexane

0.2 mM NAD(P)H

—

Classical aqueous control (in %) | Raw activity data of reaction temperature screening

Activity - Classical

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

Ethanol

5 % v/v

100.0

98.4

Direct

Continuous

50 mM sodium phosphate

7.4

40°C

10 mM Cyclohex-2-enone

Cyclohexane

0.2 mM NAD(P)H

—

Classical aqueous control (in %) | Raw activity data of reaction temperature screening

Activity - Classical

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

Ethanol

10 % v/v

100.0

34.4

Direct

Continuous

50 mM sodium phosphate

7.4

40°C

10 mM Cyclohex-2-enone

Cyclohexane

0.2 mM NAD(P)H

—

Classical aqueous control (in %) | Raw activity data of reaction temperature screening

Activity - Classical

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

Ethanol

15 % v/v

100.0

6.6

Direct

Continuous

50 mM sodium phosphate

7.4

40°C

10 mM Cyclohex-2-enone

Cyclohexane

0.2 mM NAD(P)H

—

Classical aqueous control (in %) | Raw activity data of reaction temperature screening

Activity - Classical

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

Ethanol

20 % v/v

100.0

8.2

Direct

Continuous

50 mM sodium phosphate

7.4

40°C

10 mM Cyclohex-2-enone

Cyclohexane

0.2 mM NAD(P)H

—

Classical aqueous control (in %) | Raw activity data of reaction temperature screening

Activity - Classical

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

Ethanol

25 % v/v

100.0

6.6

Direct

Continuous

50 mM sodium phosphate

7.4

40°C

10 mM Cyclohex-2-enone

Cyclohexane

0.2 mM NAD(P)H

—

Classical aqueous control (in %) | Raw activity data of reaction temperature screening

Activity - Classical

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

Ethanol

30 % v/v

100.0

6.6

Direct

Continuous

50 mM sodium phosphate

7.4

40°C

10 mM Cyclohex-2-enone

Cyclohexane

0.2 mM NAD(P)H

—

Classical aqueous control (in %) | Raw activity data of reaction temperature screening

Activity - Classical

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

Ethanol

35 % v/v

100.0

4.9

Direct

Continuous

50 mM sodium phosphate

7.4

40°C

10 mM Cyclohex-2-enone

Cyclohexane

0.2 mM NAD(P)H

—

Classical aqueous control (in %) | Raw activity data of reaction temperature screening

Activity - Classical

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

Ethanol

40 % v/v

100.0

18.0

Direct

Continuous

50 mM sodium phosphate

7.4

40°C

10 mM Cyclohex-2-enone

Cyclohexane

0.2 mM NAD(P)H

—

Classical aqueous control (in %) | Raw activity data of reaction temperature screening

Activity - Classical

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

Ethanol

45 % v/v

100.0

0.0

Direct

Continuous

50 mM sodium phosphate

7.4

40°C

10 mM Cyclohex-2-enone

Cyclohexane

0.2 mM NAD(P)H

—

Classical aqueous control (in %) | Raw activity data of reaction temperature screening

Activity - Classical

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

Ethanol

5 % v/v

100.0

69.0

Direct

Continuous

50 mM sodium phosphate

7.4

45°C

10 mM Cyclohex-2-enone

Cyclohexane

0.2 mM NAD(P)H

—

Classical aqueous control (in %) | Raw activity data of reaction temperature screening

Activity - Classical

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

Ethanol

10 % v/v

100.0

6.9

Direct

Continuous

50 mM sodium phosphate

7.4

45°C

10 mM Cyclohex-2-enone

Cyclohexane

0.2 mM NAD(P)H

—

Classical aqueous control (in %) | Raw activity data of reaction temperature screening

Activity - Classical

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

Ethanol

15 % v/v

100.0

13.8

Direct

Continuous

50 mM sodium phosphate

7.4

45°C

10 mM Cyclohex-2-enone

Cyclohexane

0.2 mM NAD(P)H

—

Classical aqueous control (in %) | Raw activity data of reaction temperature screening

Activity - Classical

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

Ethanol

20 % v/v

100.0

27.6

Direct

Continuous

50 mM sodium phosphate

7.4

45°C

10 mM Cyclohex-2-enone

Cyclohexane

0.2 mM NAD(P)H

—

Classical aqueous control (in %) | Raw activity data of reaction temperature screening

Activity - Classical

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

Ethanol

25 % v/v

100.0

13.8

Direct

Continuous

50 mM sodium phosphate

7.4

45°C

10 mM Cyclohex-2-enone

Cyclohexane

0.2 mM NAD(P)H

—

Classical aqueous control (in %) | Raw activity data of reaction temperature screening

Activity - Classical

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

Ethanol

30 % v/v

100.0

13.8

Direct

Continuous

50 mM sodium phosphate

7.4

45°C

10 mM Cyclohex-2-enone

Cyclohexane

0.2 mM NAD(P)H

—

Classical aqueous control (in %) | Raw activity data of reaction temperature screening

Activity - Classical

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

Ethanol

35 % v/v

100.0

0.0

Direct

Continuous

50 mM sodium phosphate

7.4

45°C

10 mM Cyclohex-2-enone

Cyclohexane

0.2 mM NAD(P)H

—

Classical aqueous control (in %) | Raw activity data of reaction temperature screening

Activity - Classical

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

Ethanol

40 % v/v

100.0

20.7

Direct

Continuous

50 mM sodium phosphate

7.4

45°C

10 mM Cyclohex-2-enone

Cyclohexane

0.2 mM NAD(P)H

—

Classical aqueous control (in %) | Raw activity data of reaction temperature screening

Activity - Classical

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

Ethanol

45 % v/v

100.0

6.9

Direct

Continuous

50 mM sodium phosphate

7.4

45°C

10 mM Cyclohex-2-enone

Cyclohexane

0.2 mM NAD(P)H

—

Classical aqueous control (in %) | Raw activity data of reaction temperature screening

Activity - Classical

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

Methanol

5 % v/v

100.0

93.7

Direct

Continuous

50 mM sodium phosphate

7.4

25°C

10 mM Cyclohex-2-enone

Cyclohexane

0.2 mM NAD(P)H

—

Classical aqueous control (in %) | Raw activity data of reaction temperature screening

Activity - Classical

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

Methanol

10 % v/v

100.0

115.8

Direct

Continuous

50 mM sodium phosphate

7.4

25°C

10 mM Cyclohex-2-enone

Cyclohexane

0.2 mM NAD(P)H

—

Classical aqueous control (in %) | Raw activity data of reaction temperature screening

Activity - Classical

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

Methanol

15 % v/v

100.0

106.3

Direct

Continuous

50 mM sodium phosphate

7.4

25°C

10 mM Cyclohex-2-enone

Cyclohexane

0.2 mM NAD(P)H

—

Classical aqueous control (in %) | Raw activity data of reaction temperature screening

Activity - Classical

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

Methanol

20 % v/v

100.0

96.8

Direct

Continuous

50 mM sodium phosphate

7.4

25°C

10 mM Cyclohex-2-enone

Cyclohexane

0.2 mM NAD(P)H

—

Classical aqueous control (in %) | Raw activity data of reaction temperature screening

Activity - Classical

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

Methanol

25 % v/v

100.0

63.2

Direct

Continuous

50 mM sodium phosphate

7.4

25°C

10 mM Cyclohex-2-enone

Cyclohexane

0.2 mM NAD(P)H

—

Classical aqueous control (in %) | Raw activity data of reaction temperature screening

Activity - Classical

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

Methanol

30 % v/v

100.0

20.0

Direct

Continuous

50 mM sodium phosphate

7.4

25°C

10 mM Cyclohex-2-enone

Cyclohexane

0.2 mM NAD(P)H

—

Classical aqueous control (in %) | Raw activity data of reaction temperature screening

Activity - Classical

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

Methanol

35 % v/v

100.0

11.6

Direct

Continuous

50 mM sodium phosphate

7.4

25°C

10 mM Cyclohex-2-enone

Cyclohexane

0.2 mM NAD(P)H

—

Classical aqueous control (in %) | Raw activity data of reaction temperature screening

Activity - Classical

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

Methanol

40 % v/v

100.0

12.6

Direct

Continuous

50 mM sodium phosphate

7.4

25°C

10 mM Cyclohex-2-enone

Cyclohexane

0.2 mM NAD(P)H

—

Classical aqueous control (in %) | Raw activity data of reaction temperature screening

Activity - Classical

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

Methanol

45 % v/v

100.0

15.8

Direct

Continuous

50 mM sodium phosphate

7.4

25°C

10 mM Cyclohex-2-enone

Cyclohexane

0.2 mM NAD(P)H

—

Classical aqueous control (in %) | Raw activity data of reaction temperature screening

Activity - Classical

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

Methanol

5 % v/v

100.0

103.3

Direct

Continuous

50 mM sodium phosphate

7.4

30°C

10 mM Cyclohex-2-enone

Cyclohexane

0.2 mM NAD(P)H

—

Classical aqueous control (in %) | Raw activity data of reaction temperature screening

Activity - Classical

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

Methanol

10 % v/v

100.0

120.7

Direct

Continuous

50 mM sodium phosphate

7.4

30°C

10 mM Cyclohex-2-enone

Cyclohexane

0.2 mM NAD(P)H

—

Classical aqueous control (in %) | Raw activity data of reaction temperature screening

Activity - Classical

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

Methanol

15 % v/v

100.0

112.0

Direct

Continuous

50 mM sodium phosphate

7.4

30°C

10 mM Cyclohex-2-enone

Cyclohexane

0.2 mM NAD(P)H

—

Classical aqueous control (in %) | Raw activity data of reaction temperature screening

Activity - Classical

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

Methanol

20 % v/v

100.0

90.2

Direct

Continuous

50 mM sodium phosphate

7.4

30°C

10 mM Cyclohex-2-enone

Cyclohexane

0.2 mM NAD(P)H

—

Classical aqueous control (in %) | Raw activity data of reaction temperature screening

Activity - Classical

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

Methanol

25 % v/v

100.0

42.4

Direct

Continuous

50 mM sodium phosphate

7.4

30°C

10 mM Cyclohex-2-enone

Cyclohexane

0.2 mM NAD(P)H

—

Classical aqueous control (in %) | Raw activity data of reaction temperature screening

Activity - Classical

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

Methanol

30 % v/v

100.0

9.8

Direct

Continuous

50 mM sodium phosphate

7.4

30°C

10 mM Cyclohex-2-enone

Cyclohexane

0.2 mM NAD(P)H

—

Classical aqueous control (in %) | Raw activity data of reaction temperature screening

Activity - Classical

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

Methanol

35 % v/v

100.0

0.0

Direct

Continuous

50 mM sodium phosphate

7.4

30°C

10 mM Cyclohex-2-enone

Cyclohexane

0.2 mM NAD(P)H

—

Classical aqueous control (in %) | Raw activity data of reaction temperature screening

Activity - Classical

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

Methanol

40 % v/v

100.0

3.3

Direct

Continuous

50 mM sodium phosphate

7.4

30°C

10 mM Cyclohex-2-enone

Cyclohexane

0.2 mM NAD(P)H

—

Classical aqueous control (in %) | Raw activity data of reaction temperature screening

Activity - Classical

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

Methanol

45 % v/v

100.0

3.3

Direct

Continuous

50 mM sodium phosphate

7.4

30°C

10 mM Cyclohex-2-enone

Cyclohexane

0.2 mM NAD(P)H

—

Classical aqueous control (in %) | Raw activity data of reaction temperature screening

Activity - Classical

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

Methanol

5 % v/v

100.0

102.1

Direct

Continuous

50 mM sodium phosphate

7.4

35°C

10 mM Cyclohex-2-enone

Cyclohexane

0.2 mM NAD(P)H

—

Classical aqueous control (in %) | Raw activity data of reaction temperature screening

Activity - Classical

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

Methanol

10 % v/v

100.0

101.0

Direct

Continuous

50 mM sodium phosphate

7.4

35°C

10 mM Cyclohex-2-enone

Cyclohexane

0.2 mM NAD(P)H

—

Classical aqueous control (in %) | Raw activity data of reaction temperature screening

Activity - Classical

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

Methanol

15 % v/v

100.0

91.8

Direct

Continuous

50 mM sodium phosphate

7.4

35°C

10 mM Cyclohex-2-enone

Cyclohexane

0.2 mM NAD(P)H

—

Classical aqueous control (in %) | Raw activity data of reaction temperature screening

Activity - Classical

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

Methanol

20 % v/v

100.0

33.0

Direct

Continuous

50 mM sodium phosphate

7.4

35°C

10 mM Cyclohex-2-enone

Cyclohexane

0.2 mM NAD(P)H

—

Classical aqueous control (in %) | Raw activity data of reaction temperature screening

Activity - Classical

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

Methanol

25 % v/v

100.0

7.2

Direct

Continuous

50 mM sodium phosphate

7.4

35°C

10 mM Cyclohex-2-enone

Cyclohexane

0.2 mM NAD(P)H

—

Classical aqueous control (in %) | Raw activity data of reaction temperature screening

Activity - Classical

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

Methanol

30 % v/v

100.0

5.2

Direct

Continuous

50 mM sodium phosphate

7.4

35°C

10 mM Cyclohex-2-enone

Cyclohexane

0.2 mM NAD(P)H

—

Classical aqueous control (in %) | Raw activity data of reaction temperature screening

Activity - Classical

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

Methanol

35 % v/v

100.0

2.1

Direct

Continuous

50 mM sodium phosphate

7.4

35°C

10 mM Cyclohex-2-enone

Cyclohexane

0.2 mM NAD(P)H

—

Classical aqueous control (in %) | Raw activity data of reaction temperature screening

Activity - Classical

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

Methanol

40 % v/v

100.0

7.2

Direct

Continuous

50 mM sodium phosphate

7.4

35°C

10 mM Cyclohex-2-enone

Cyclohexane

0.2 mM NAD(P)H

—

Classical aqueous control (in %) | Raw activity data of reaction temperature screening

Activity - Classical

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

Methanol

45 % v/v

100.0

3.1

Direct

Continuous

50 mM sodium phosphate

7.4

35°C

10 mM Cyclohex-2-enone

Cyclohexane

0.2 mM NAD(P)H

—

Classical aqueous control (in %) | Raw activity data of reaction temperature screening

Activity - Classical

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

Methanol

5 % v/v

100.0

101.5

Direct

Continuous

50 mM sodium phosphate

7.4

40°C

10 mM Cyclohex-2-enone

Cyclohexane

0.2 mM NAD(P)H

—

Classical aqueous control (in %) | Raw activity data of reaction temperature screening

Activity - Classical

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

Methanol

10 % v/v

100.0

61.2

Direct

Continuous

50 mM sodium phosphate

7.4

40°C

10 mM Cyclohex-2-enone

Cyclohexane

0.2 mM NAD(P)H

—

Classical aqueous control (in %) | Raw activity data of reaction temperature screening

Activity - Classical

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

Methanol

15 % v/v

100.0

29.9

Direct

Continuous

50 mM sodium phosphate

7.4

40°C

10 mM Cyclohex-2-enone

Cyclohexane

0.2 mM NAD(P)H

—

Classical aqueous control (in %) | Raw activity data of reaction temperature screening

Activity - Classical

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

Methanol

20 % v/v

100.0

7.5

Direct

Continuous

50 mM sodium phosphate

7.4

40°C

10 mM Cyclohex-2-enone

Cyclohexane

0.2 mM NAD(P)H

—

Classical aqueous control (in %) | Raw activity data of reaction temperature screening

Activity - Classical

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

Methanol

25 % v/v

100.0

3.0

Direct

Continuous

50 mM sodium phosphate

7.4

40°C

10 mM Cyclohex-2-enone

Cyclohexane

0.2 mM NAD(P)H

—

Classical aqueous control (in %) | Raw activity data of reaction temperature screening

Activity - Classical

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

Methanol

30 % v/v

100.0

4.5

Direct

Continuous

50 mM sodium phosphate

7.4

40°C

10 mM Cyclohex-2-enone

Cyclohexane

0.2 mM NAD(P)H

—

Classical aqueous control (in %) | Raw activity data of reaction temperature screening

Activity - Classical

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

Methanol

35 % v/v

100.0

4.5

Direct

Continuous

50 mM sodium phosphate

7.4

40°C

10 mM Cyclohex-2-enone

Cyclohexane

0.2 mM NAD(P)H

—

Classical aqueous control (in %) | Raw activity data of reaction temperature screening

Activity - Classical

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

Methanol

40 % v/v

100.0

7.5

Direct

Continuous

50 mM sodium phosphate

7.4

40°C

10 mM Cyclohex-2-enone

Cyclohexane

0.2 mM NAD(P)H

—

Classical aqueous control (in %) | Raw activity data of reaction temperature screening

Activity - Classical

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

Methanol

45 % v/v

100.0

3.0

Direct

Continuous

50 mM sodium phosphate

7.4

40°C

10 mM Cyclohex-2-enone

Cyclohexane

0.2 mM NAD(P)H

—

Classical aqueous control (in %) | Raw activity data of reaction temperature screening

Activity - Classical

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

Methanol

5 % v/v

100.0

80.0

Direct

Continuous

50 mM sodium phosphate

7.4

45°C

10 mM Cyclohex-2-enone

Cyclohexane

0.2 mM NAD(P)H

—

Classical aqueous control (in %) | Raw activity data of reaction temperature screening

Activity - Classical

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

Methanol

10 % v/v

100.0

34.3

Direct

Continuous

50 mM sodium phosphate

7.4

45°C

10 mM Cyclohex-2-enone

Cyclohexane

0.2 mM NAD(P)H

—

Classical aqueous control (in %) | Raw activity data of reaction temperature screening

Activity - Classical

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

Methanol

15 % v/v

100.0

2.9

Direct

Continuous

50 mM sodium phosphate

7.4

45°C

10 mM Cyclohex-2-enone

Cyclohexane

0.2 mM NAD(P)H

—

Classical aqueous control (in %) | Raw activity data of reaction temperature screening

Activity - Classical

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

Methanol

20 % v/v

100.0

8.6

Direct

Continuous

50 mM sodium phosphate

7.4

45°C

10 mM Cyclohex-2-enone

Cyclohexane

0.2 mM NAD(P)H

—

Classical aqueous control (in %) | Raw activity data of reaction temperature screening

Activity - Classical

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

Methanol

25 % v/v

100.0

8.6

Direct

Continuous

50 mM sodium phosphate

7.4

45°C

10 mM Cyclohex-2-enone

Cyclohexane

0.2 mM NAD(P)H

—

Classical aqueous control (in %) | Raw activity data of reaction temperature screening

Activity - Classical

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

Methanol

30 % v/v

100.0

8.6

Direct

Continuous

50 mM sodium phosphate

7.4

45°C

10 mM Cyclohex-2-enone

Cyclohexane

0.2 mM NAD(P)H

—

Classical aqueous control (in %) | Raw activity data of reaction temperature screening

Activity - Classical

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

Methanol

35 % v/v

100.0

11.4

Direct

Continuous

50 mM sodium phosphate

7.4

45°C

10 mM Cyclohex-2-enone

Cyclohexane

0.2 mM NAD(P)H

—

Classical aqueous control (in %) | Raw activity data of reaction temperature screening

Activity - Classical

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

Methanol

40 % v/v

100.0

17.1

Direct

Continuous

50 mM sodium phosphate

7.4

45°C

10 mM Cyclohex-2-enone

Cyclohexane

0.2 mM NAD(P)H

—

Classical aqueous control (in %) | Raw activity data of reaction temperature screening

Activity - Classical

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

Methanol

45 % v/v

100.0

8.6

Direct

Continuous

50 mM sodium phosphate

7.4

45°C

10 mM Cyclohex-2-enone

Cyclohexane

0.2 mM NAD(P)H

—

Classical aqueous control (in %) | Raw activity data of reaction temperature screening

Stability - C50

Volume of organic solvent that leads to half-unfolding, measured by nanoDSF

Dimethylformamide (DMF)

—

26.0

% (v/v)

Direct

Continuous

50 mM sodium phosphate

7.4

25°C

—

Not applicable control (in % (v/v))

Stability - C50

Volume of organic solvent that leads to half-unfolding, measured by nanoDSF

Dimethylformamide (DMF)

—

23.0

% (v/v)

Direct

Continuous

50 mM sodium phosphate

7.4

30°C

—

Not applicable control (in % (v/v))

Stability - C50

Volume of organic solvent that leads to half-unfolding, measured by nanoDSF

Dimethylformamide (DMF)

—

19.0

% (v/v)

Direct

Continuous

50 mM sodium phosphate

7.4

35°C

—

Not applicable control (in % (v/v))

Stability - C50

Volume of organic solvent that leads to half-unfolding, measured by nanoDSF

Dimethylformamide (DMF)

—

15.0

% (v/v)

Direct

Continuous

50 mM sodium phosphate

7.4

40°C

—

Not applicable control (in % (v/v))

Stability - C50

Volume of organic solvent that leads to half-unfolding, measured by nanoDSF

Dimethylformamide (DMF)

—

11.0

% (v/v)

Direct

Continuous

50 mM sodium phosphate

7.4

45°C

—

Not applicable control (in % (v/v))

Stability - C50

Volume of organic solvent that leads to half-unfolding, measured by nanoDSF

Dimethylformamide (DMF)

—

9.0

% (v/v)

Direct

Continuous

50 mM sodium phosphate

7.4

50°C

—

Not applicable control (in % (v/v))

Stability - C50

Volume of organic solvent that leads to half-unfolding, measured by nanoDSF

Dimethyl Sulfoxide (DMSO)

—

33.0

% (v/v)

Direct

Continuous

50 mM sodium phosphate

7.4

30°C

—

Not applicable control (in % (v/v))

Stability - C50

Volume of organic solvent that leads to half-unfolding, measured by nanoDSF

Dimethyl Sulfoxide (DMSO)

—

26.0

% (v/v)

Direct

Continuous

50 mM sodium phosphate

7.4

35°C

—

Not applicable control (in % (v/v))

Stability - C50

Volume of organic solvent that leads to half-unfolding, measured by nanoDSF

Dimethyl Sulfoxide (DMSO)

—

18.0

% (v/v)

Direct

Continuous

50 mM sodium phosphate

7.4

40°C

—

Not applicable control (in % (v/v))

Stability - C50

Volume of organic solvent that leads to half-unfolding, measured by nanoDSF

Dimethyl Sulfoxide (DMSO)

—

11.0

% (v/v)

Direct

Continuous

50 mM sodium phosphate

7.4

45°C

—

Not applicable control (in % (v/v))

Stability - C50

Volume of organic solvent that leads to half-unfolding, measured by nanoDSF

1-Propanol

—

16.0

% (v/v)

Direct

Continuous

50 mM sodium phosphate

7.4

25°C

—

Not applicable control (in % (v/v))

Stability - C50

Volume of organic solvent that leads to half-unfolding, measured by nanoDSF

1-Propanol

—

14.0

% (v/v)

Direct

Continuous

50 mM sodium phosphate

7.4

30°C

—

Not applicable control (in % (v/v))

Stability - C50

Volume of organic solvent that leads to half-unfolding, measured by nanoDSF

1-Propanol

—

12.0

% (v/v)

Direct

Continuous

50 mM sodium phosphate

7.4

35°C

—

Not applicable control (in % (v/v))

Stability - C50

Volume of organic solvent that leads to half-unfolding, measured by nanoDSF

1-Propanol

—

9.0

% (v/v)

Direct

Continuous

50 mM sodium phosphate

7.4

40°C

—

Not applicable control (in % (v/v))

Stability - Tm

Melting temperature measured by ThermoFMN in the presence of organic solvent

Dimethyl Sulfoxide (DMSO)

5 % v/v

58.0

55.0

°C

Direct

Continuous

50 mM sodium phosphate

7.4

—

Classical aqueous control (in °C)

Stability - Tm

Melting temperature measured by ThermoFMN in the presence of organic solvent

Dimethyl Sulfoxide (DMSO)

10 % v/v

58.0

53.33

°C

Direct

Continuous

50 mM sodium phosphate

7.4

—

Classical aqueous control (in °C)

Stability - Tm

Melting temperature measured by ThermoFMN in the presence of organic solvent

Dimethyl Sulfoxide (DMSO)

15 % v/v

58.0

53.0

°C

Direct

Continuous

50 mM sodium phosphate

7.4

—

Classical aqueous control (in °C)

Stability - Tm

Melting temperature measured by ThermoFMN in the presence of organic solvent

Dimethyl Sulfoxide (DMSO)

20 % v/v

58.0

52.0

°C

Direct

Continuous

50 mM sodium phosphate

7.4

—

Classical aqueous control (in °C)

Stability - Tm

Melting temperature measured by ThermoFMN in the presence of organic solvent

Dimethyl Sulfoxide (DMSO)

25 % v/v

58.0

51.67

°C

Direct

Continuous

50 mM sodium phosphate

7.4

—

Classical aqueous control (in °C)

Stability - Tm

Melting temperature measured by ThermoFMN in the presence of organic solvent

Dimethyl Sulfoxide (DMSO)

30 % v/v

58.0

52.0

°C

Direct

Continuous

50 mM sodium phosphate

7.4

—

Classical aqueous control (in °C)

Stability - Tm

Melting temperature measured by ThermoFMN in the presence of organic solvent

Ethanol

5 % v/v

58.0

53.0

°C

Direct

Continuous

50 mM sodium phosphate

7.4

—

Classical aqueous control (in °C)

Stability - Tm

Melting temperature measured by ThermoFMN in the presence of organic solvent

Ethanol

10 % v/v

58.0

46.33

°C

Direct

Continuous

50 mM sodium phosphate

7.4

—

Classical aqueous control (in °C)

Stability - Tm

Melting temperature measured by ThermoFMN in the presence of organic solvent

Ethanol

15 % v/v

58.0

42.67

°C

Direct

Continuous

50 mM sodium phosphate

7.4

—

Classical aqueous control (in °C)

Stability - Tm

Melting temperature measured by ThermoFMN in the presence of organic solvent

Ethanol

20 % v/v

58.0

40.67

°C

Direct

Continuous

50 mM sodium phosphate

7.4

—

Classical aqueous control (in °C)

Stability - Tm

Melting temperature measured by ThermoFMN in the presence of organic solvent

Ethanol

25 % v/v

58.0

34.33

°C

Direct

Continuous

50 mM sodium phosphate

7.4

—

Classical aqueous control (in °C)

Stability - Tm

Melting temperature measured by ThermoFMN in the presence of organic solvent

Ethanol

30 % v/v

58.0

35.67

°C

Direct

Continuous

50 mM sodium phosphate

7.4

—

Classical aqueous control (in °C)

Stability - Tm

Melting temperature measured by ThermoFMN in the presence of organic solvent

Methanol

5 % v/v

58.0

57.0

°C

Direct

Continuous

50 mM sodium phosphate

7.4

—

Classical aqueous control (in °C)

Stability - Tm

Melting temperature measured by ThermoFMN in the presence of organic solvent

Methanol

10 % v/v

58.0

52.33

°C

Direct

Continuous

50 mM sodium phosphate

7.4

—

Classical aqueous control (in °C)

Stability - Tm

Melting temperature measured by ThermoFMN in the presence of organic solvent

Methanol

15 % v/v

58.0

49.67

°C

Direct

Continuous

50 mM sodium phosphate

7.4

—

Classical aqueous control (in °C)

Stability - Tm

Melting temperature measured by ThermoFMN in the presence of organic solvent

Methanol

20 % v/v

58.0

45.0

°C

Direct

Continuous

50 mM sodium phosphate

7.4

—

Classical aqueous control (in °C)

Stability - Tm

Melting temperature measured by ThermoFMN in the presence of organic solvent

Methanol

25 % v/v

58.0

41.0

°C

Direct

Continuous

50 mM sodium phosphate

7.4

—

Classical aqueous control (in °C)

Stability - Tm

Melting temperature measured by ThermoFMN in the presence of organic solvent

Methanol

30 % v/v

58.0

38.33

°C

Direct

Continuous

50 mM sodium phosphate

7.4

—

Classical aqueous control (in °C)

Stability - Tm

Melting temperature measured by ThermoFMN in the presence of organic solvent

Isopropanol

5 % v/v

58.0

49.0

°C

Direct

Continuous

50 mM sodium phosphate

7.4

—

Classical aqueous control (in °C)

Stability - Tm

Melting temperature measured by ThermoFMN in the presence of organic solvent

Isopropanol

10 % v/v

58.0

43.67

°C

Direct

Continuous

50 mM sodium phosphate

7.4

—

Classical aqueous control (in °C)

Stability - Tm

Melting temperature measured by ThermoFMN in the presence of organic solvent

Isopropanol

15 % v/v

58.0

38.33

°C

Direct

Continuous

50 mM sodium phosphate

7.4

—

Classical aqueous control (in °C)

Stability - Tm

Melting temperature measured by ThermoFMN in the presence of organic solvent

Isopropanol

20 % v/v

58.0

35.67

°C

Direct

Continuous

50 mM sodium phosphate

7.4

—

Classical aqueous control (in °C)

Stability - Tm

Melting temperature measured by ThermoFMN in the presence of organic solvent

Isopropanol

25 % v/v

58.0

32.67

°C

Direct

Continuous

50 mM sodium phosphate

7.4

—

Classical aqueous control (in °C)

Stability - Tm

Melting temperature measured by ThermoFMN in the presence of organic solvent

Isopropanol

30 % v/v

58.0

28.67

°C

Direct

Continuous

50 mM sodium phosphate

7.4

—

Classical aqueous control (in °C)

Stability - Tm

Melting temperature measured by ThermoFMN in the presence of organic solvent

1-Propanol

5 % v/v

58.0

47.0

°C

Direct

Continuous

50 mM sodium phosphate

7.4

—

Classical aqueous control (in °C)

Stability - Tm

Melting temperature measured by ThermoFMN in the presence of organic solvent

1-Propanol

10 % v/v

58.0

36.0

°C

Direct

Continuous

50 mM sodium phosphate

7.4

—

Classical aqueous control (in °C)

Stability - Tm

Melting temperature measured by ThermoFMN in the presence of organic solvent

1-Propanol

15 % v/v

58.0

31.33

°C

Direct

Continuous

50 mM sodium phosphate

7.4

—

Classical aqueous control (in °C)

Stability - Tm

Melting temperature measured by ThermoFMN in the presence of organic solvent

1-Propanol

20 % v/v

58.0

31.0

°C

Direct

Continuous

50 mM sodium phosphate

7.4

—

Classical aqueous control (in °C)

Stability - Tm

Melting temperature measured by ThermoFMN in the presence of organic solvent

1-Propanol

25 % v/v

58.0

28.67

°C

Direct

Continuous

50 mM sodium phosphate

7.4

—

Classical aqueous control (in °C)

Stability - Tm

Melting temperature measured by ThermoFMN in the presence of organic solvent

1-Propanol

30 % v/v

58.0

27.0

°C

Direct

Continuous

50 mM sodium phosphate

7.4

—

Classical aqueous control (in °C)

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Visualization : Activity — Classical

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

Visualization : Stability — T_m

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

Visualization : Stability — C₅₀

One bar per measurement. Colour = solvent, shade = temperature. Hover for details.

Structure

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Ene-reductase DrER from Deinococcus radiodurans

Enzyme Description

Sequence

Experimental Data (164 measurements)

Visualization : Activity — Classical

Visualization : Stability — Tm

Visualization : Stability — C50

Structure

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Visualization : Stability — T_m

Visualization : Stability — C₅₀