Ene-reductase OYE1 from Saccharomyces pastorianus

Enzyme Description

Species

Extremophile

EC Number

1.6.99.1 ↗

Sequence

Length: 400 amino acids

MSFVKDFKPQALGDTNLFKPIKIGNNELLHRAVIPPLTRMRALHPGNIPNRDWAVEYYTQRAQRPGTMIITEGAFISPQAGGYDNAPGVWSEEQMVEWTKIFNAIHEKKSFVWVQLWVLGWAAFPDNLARDGLRYDSASDNVFMDAEQEAKAKKANNPQHSLTKDEIKQYIKEYVQAAKNSIAAGADGVEIHSANGYLLNQFLDPHSNTRTDEYGGSIENRARFTLEVVDALVEAIGHEKVGLRLSPYGVFNSMSGGAETGIVAQYAYVAGELEKRAKAGKRLAFVHLVEPRVTNPFLTEGEGEYEGGSNDFVYSIWKGPVIRAGNFALHPEVVREEVKDKRTLIGYGRFFISNPDLVDRLEKGLPLNKYDRDTFYQMSAHGYIDYPTYEEALKLGWDKK

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 - Tm

150 measurements ▼

Database ID

UniProt: Q02899 ↗

Sequence Annotation

Explicit - Provided UniProt Accession Number

Protein Source

Recombinant, host bacterium Escherichia coli BL21 (DE3)

Experimental Data (150 measurements)

150 measurements — page 1 of 8

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	109.2	%	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	Dimethyl Sulfoxide (DMSO)	10 % v/v	100.0	103.1	%	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	100.0	%	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)	20 % v/v	100.0	105.3	%	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)	25 % v/v	100.0	100.8	%	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)	30 % v/v	100.0	100.8	%	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	Ethanol	5 % v/v	100.0	106.0	%	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	119.5	%	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	15 % v/v	100.0	132.2	%	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
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	134.2	%	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
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	140.9	%	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	Ethanol	30 % v/v	100.0	165.1	%	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	5 % v/v	100.0	111.0	%	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	10 % v/v	100.0	116.8	%	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	Methanol	15 % v/v	100.0	133.5	%	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	20 % v/v	100.0	143.9	%	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	25 % v/v	100.0	156.1	%	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	166.5	%	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	126.2	%	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	139.3	%	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)

5 % v/v

100.0

109.2

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

Dimethyl Sulfoxide (DMSO)

10 % v/v

100.0

103.1

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

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

105.3

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)

25 % v/v

100.0

100.8

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)

30 % v/v

100.0

100.8

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

Ethanol

5 % v/v

100.0

106.0

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

119.5

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

15 % v/v

100.0

132.2

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

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

134.2

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

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

140.9

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

Ethanol

30 % v/v

100.0

165.1

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

5 % v/v

100.0

111.0

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

10 % v/v

100.0

116.8

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

Methanol

15 % v/v

100.0

133.5

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

20 % v/v

100.0

143.9

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

25 % v/v

100.0

156.1

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

166.5

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

126.2

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

139.3

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

152.5

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

Isopropanol

20 % v/v

100.0

147.5

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

Isopropanol

25 % v/v

100.0

163.9

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

168.0

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

5 % v/v

100.0

122.3

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

10 % v/v

100.0

140.8

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

139.2

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

1-Propanol

20 % v/v

100.0

143.1

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

1-Propanol

25 % v/v

100.0

141.5

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

30 % v/v

100.0

150.8

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

Ethanol

5 % v/v

100.0

140.0

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

10 % v/v

100.0

135.0

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

15 % v/v

100.0

160.0

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

180.0

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

25 % v/v

100.0

135.0

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

75.0

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

35.0

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

40 % v/v

100.0

10.0

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

45 % v/v

100.0

5.0

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

5 % v/v

100.0

139.1

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

160.9

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

15 % v/v

100.0

156.5

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

20 % v/v

100.0

152.2

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

Ethanol

25 % v/v

100.0

91.3

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

30 % v/v

100.0

65.2

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

Ethanol

35 % v/v

100.0

13.0

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

8.7

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

45 % v/v

100.0

17.4

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

130.6

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

10 % v/v

100.0

144.4

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

144.4

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

20 % v/v

100.0

88.9

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

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

30 % v/v

100.0

5.6

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

35 % v/v

100.0

2.8

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

40 % v/v

100.0

2.8

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

45 % v/v

100.0

5.6

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

5 % v/v

100.0

103.0

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

10 % v/v

100.0

63.6

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

15 % v/v

100.0

27.3

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

20 % v/v

100.0

6.1

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

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

Ethanol

30 % 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

Ethanol

35 % 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

Ethanol

40 % 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

Ethanol

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

Ethanol

5 % v/v

100.0

170.0

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

120.0

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

15 % v/v

100.0

50.0

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

20 % v/v

100.0

20.0

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

25 % v/v

100.0

10.0

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

30 % v/v

100.0

20.0

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

35 % v/v

100.0

30.0

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

10.0

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

45 % v/v

100.0

20.0

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

5 % v/v

100.0

87.5

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

10 % v/v

100.0

125.0

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

15 % v/v

100.0

137.5

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

154.2

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

25 % v/v

100.0

129.2

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

79.2

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

35 % v/v

100.0

12.5

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

40 % v/v

100.0

8.3

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

4.2

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

5 % v/v

100.0

138.1

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

171.4

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

190.5

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

Methanol

20 % v/v

100.0

204.8

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

25 % v/v

100.0

142.9

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

30 % v/v

100.0

66.7

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

35 % v/v

100.0

19.0

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

14.3

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

14.3

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

124.3

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

Methanol

10 % v/v

100.0

140.5

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

Methanol

15 % v/v

100.0

148.6

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

Methanol

20 % v/v

100.0

108.1

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

25 % v/v

100.0

16.2

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

2.7

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

Methanol

35 % v/v

100.0

2.7

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

Methanol

40 % 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

Methanol

45 % v/v

100.0

2.7

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

Methanol

5 % v/v

100.0

108.7

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

10 % v/v

100.0

73.9

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

15 % v/v

100.0

39.1

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

8.7

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

25 % v/v

100.0

8.7

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

4.3

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

35 % 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

Methanol

40 % v/v

100.0

4.3

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

45 % v/v

100.0

4.3

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

5 % v/v

100.0

171.4

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

10 % v/v

100.0

114.3

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

57.1

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

20 % v/v

100.0

14.3

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

25 % v/v

100.0

28.6

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

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

Methanol

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

Methanol

40 % v/v

100.0

14.3

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

45 % 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

Stability - Tm

Melting temperature measured by ThermoFMN in the presence of organic solvent

Dimethyl Sulfoxide (DMSO)

5 % v/v

45.3

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

Dimethyl Sulfoxide (DMSO)

10 % v/v

45.3

43.33

°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

45.3

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

Dimethyl Sulfoxide (DMSO)

20 % v/v

45.3

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

Dimethyl Sulfoxide (DMSO)

25 % v/v

45.3

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

Dimethyl Sulfoxide (DMSO)

30 % v/v

45.3

37.33

°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

5 % v/v

45.3

43.33

°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

10 % v/v

45.3

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

15 % v/v

45.3

39.67

°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

45.3

37.67

°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

45.3

35.33

°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

45.3

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

Methanol

5 % v/v

45.3

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

10 % v/v

45.3

43.0

°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

15 % v/v

45.3

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

Methanol

20 % v/v

45.3

40.33

°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

45.3

37.67

°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

30 % v/v

45.3

34.67

°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

5 % v/v

45.3

44.0

°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

45.3

41.33

°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

45.3

37.33

°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

45.3

34.67

°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

45.3

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

30 % v/v

45.3

28.33

°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

5 % v/v

45.3

40.33

°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

10 % v/v

45.3

34.67

°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

15 % v/v

45.3

31.67

°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

20 % v/v

45.3

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

45.3

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

30 % v/v

45.3

27.33

°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.

Structure

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Ene-reductase OYE1 from Saccharomyces pastorianus

Enzyme Description

Sequence

Experimental Data (150 measurements)

Visualization : Activity — Classical

Visualization : Stability — Tm

Structure

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Visualization : Stability — T_m