Ene-reductase YqiG from Bacillus subtilis

Enzyme Description

Species

Extremophile

EC Number

1.6.99.1 ↗

Sequence

Length: 372 amino acids

MNPKYKPLFEPFTFKSGVTINNRIAVAPMTHYASNEDGTISEAELDYIIPRSKEMGMVITACANVTPDGKAFPGQPAIHDDSNIPGLKKLAQAIQAQGAKAVVQIHHGGIECPSELVPQQDVVGPSDVFDNGKQIARALTEEEVENIVKAFGEATRRAIEAGFDGVEIHGANGYLIQQFYSPKTNQRTDRWGGSDEKRLAFPLAIVDEVKKAASEHAKGAFLVGYRLSPEEPETPGLTMTETYTLVDALGDKELDYLHISLMDVNSKARRGADPTRTRMDLLNERVGNKVPLIAVGSIHSADDALAVIENGIPLVAMGREILVDPNWTVKVKEGREKQIETVIKGTDKEKYHLPEPLWQAIVNTQGWVPYKD

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

140 measurements ▼

Database ID

UniProt: P54524 ↗

Sequence Annotation

Explicit - Provided UniProt Accession Number

Protein Source

Recombinant, host bacterium Escherichia coli BL21 (DE3)

Experimental Data (140 measurements)

140 measurements — page 7 of 7

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
Stability - Tm	Melting temperature measured by ThermoFMN in the presence of organic solvent	Ethanol	25 % v/v	54.0	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	Ethanol	30 % v/v	54.0	36.33	°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	5 % v/v	54.0	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	10 % v/v	54.0	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	15 % v/v	54.0	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	Methanol	20 % v/v	54.0	44.33	°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	25 % v/v	54.0	42.33	°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	30 % v/v	54.0	40.0	°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	5 % v/v	54.0	46.0	°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	54.0	39.33	°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	15 % v/v	54.0	39.33	°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	54.0	33.67	°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	54.0	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	54.0	33.67	°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	54.0	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	1-Propanol	10 % v/v	54.0	33.0	°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	54.0	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	54.0	30.33	°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	25 % v/v	54.0	29.33	°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	30 % v/v	54.0	30.33	°C	Direct	Continuous	50 mM sodium phosphate	7.4	—	—	—	—	—	Classical aqueous control (in °C)

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

107.6

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

106.1

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)

15 % v/v

100.0

107.8

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

Dimethyl Sulfoxide (DMSO)

20 % v/v

100.0

102.7

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)

25 % v/v

100.0

106.4

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)

30 % v/v

100.0

102.7

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

103.0

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

108.6

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

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

116.4

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

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

108.4

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

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

115.5

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

Ethanol

30 % v/v

100.0

104.0

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

5 % v/v

100.0

107.5

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

98.4

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

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

93.9

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

20 % v/v

100.0

88.4

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

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

89.1

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

Methanol

30 % v/v

100.0

74.8

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

Isopropanol

5 % v/v

100.0

104.7

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

10 % v/v

100.0

106.3

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

Isopropanol

15 % v/v

100.0

108.0

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

Isopropanol

20 % v/v

100.0

107.3

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

Isopropanol

25 % v/v

100.0

101.0

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

Isopropanol

30 % v/v

100.0

91.5

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

5 % v/v

100.0

99.0

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

10 % v/v

100.0

91.8

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

15 % v/v

100.0

86.7

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

20 % v/v

100.0

77.5

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

25 % v/v

100.0

70.3

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

1-Propanol

30 % v/v

100.0

56.0

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

Ethanol

5 % v/v

100.0

96.3

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

10 % v/v

100.0

118.8

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

120.0

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

20 % v/v

100.0

123.8

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

25 % v/v

100.0

110.0

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

30 % 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 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

56.3

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

40 % v/v

100.0

28.8

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

45 % v/v

100.0

13.8

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

5 % v/v

100.0

117.9

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

10 % v/v

100.0

119.6

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

151.8

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

20 % v/v

100.0

133.9

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

25 % v/v

100.0

112.5

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

30 % v/v

100.0

78.6

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

44.6

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

17.9

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

10.7

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

5 % v/v

100.0

112.5

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

116.7

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

15 % v/v

100.0

87.5

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

20 % v/v

100.0

70.8

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

Ethanol

25 % v/v

100.0

33.3

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

16.7

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

35 % v/v

100.0

8.3

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

16.7

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

45 % v/v

100.0

0.0

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

Ethanol

5 % v/v

100.0

33.3

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

26.7

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

20.0

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

20 % v/v

100.0

20.0

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

25 % v/v

100.0

20.0

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

30 % v/v

100.0

26.7

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

26.7

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

13.3

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

26.7

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

92.4

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

Methanol

10 % v/v

100.0

104.5

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 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

98.5

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 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

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

90.9

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 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

83.3

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

35 % v/v

100.0

66.7

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

40 % v/v

100.0

30.3

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

45 % v/v

100.0

6.1

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

Methanol

5 % v/v

100.0

82.4

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

10 % v/v

100.0

80.4

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

15 % v/v

100.0

84.3

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

20 % v/v

100.0

78.4

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

25 % v/v

100.0

78.4

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

54.9

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

Methanol

35 % v/v

100.0

41.2

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

40 % v/v

100.0

21.6

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

7.8

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

5 % v/v

100.0

80.0

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

10 % v/v

100.0

56.0

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

40.0

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

20 % v/v

100.0

40.0

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

20.0

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

30 % v/v

100.0

16.0

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

35 % v/v

100.0

8.0

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

40 % v/v

100.0

28.0

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

45 % v/v

100.0

16.0

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

5 % v/v

100.0

75.0

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

10 % v/v

100.0

33.3

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

16.7

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

25.0

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

25 % v/v

100.0

16.7

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

25.0

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

35 % v/v

100.0

16.7

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

25.0

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

45 % v/v

100.0

41.7

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

Stability - C50

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

Dimethylformamide (DMF)

—

35.0

% (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)

—

14.0

% (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

1-Propanol

—

18.0

% (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

—

17.0

% (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

—

14.0

% (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 - C50

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

1-Propanol

—

6.0

% (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

—

3.0

% (v/v)

Direct

Continuous

50 mM sodium phosphate

7.4

50°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

54.0

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)

10 % v/v

54.0

51.0

°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)

15 % v/v

54.0

50.0

°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

54.0

50.0

°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

54.0

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

Dimethyl Sulfoxide (DMSO)

30 % v/v

54.0

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

Ethanol

5 % v/v

54.0

51.0

°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

54.0

45.33

°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

54.0

42.0

°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

20 % v/v

54.0

38.67

°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

54.0

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

Ethanol

30 % v/v

54.0

36.33

°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

5 % v/v

54.0

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

10 % v/v

54.0

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

15 % v/v

54.0

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

Methanol

20 % v/v

54.0

44.33

°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

25 % v/v

54.0

42.33

°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

30 % v/v

54.0

40.0

°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

5 % v/v

54.0

46.0

°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

54.0

39.33

°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

15 % v/v

54.0

39.33

°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

54.0

33.67

°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

54.0

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

54.0

33.67

°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

54.0

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

1-Propanol

10 % v/v

54.0

33.0

°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

54.0

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

54.0

30.33

°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

25 % v/v

54.0

29.33

°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

30 % v/v

54.0

30.33

°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 YqiG from Bacillus subtilis

Enzyme Description

Sequence

Experimental Data (140 measurements)

Visualization : Activity — Classical

Visualization : Stability — Tm

Visualization : Stability — C50

Structure

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Visualization : Stability — T_m

Visualization : Stability — C₅₀