Ene-reductase NCR from Zymomonas mobilis

Enzyme Description

Species

Extremophile

EC Number

1.6.99.1 ↗

Sequence

Length: 358 amino acids

MPSLFDPIRFGAFTAKNRIWMAPLTRGRATRDHVPTEIMAEYYAQRASAGLIISEATGISQEGLGWPYAPGIWSDAQVEAWLPITQAVHDAGGLIFAQLWHMGRMVPSNVSGMQPVAPSASQAPGLGHTYDGKKPYDVARALRLDEIPRLLDDYEKAARHALKAGFDGVQIHAANGYLIDEFIRDSTNHRHDEYGGAVENRIRLLKDVTERVIATIGKERTAVRLSPNGEIQGTVDSHPEQVFIPAAKMLSDLDIAFLGMREGAVDGTFGKTDQPKLSPEIRKVFKPPLVLNQDYTFETAQAALDSGVADAISFGRPFIGNPDLPRRFFEKAPLTKDVIETWYTQTPKGYTDYPLLGD

Sabrina Reich et al. (2014) — Variations in the stability of NCR ene reductase by rational enzyme loop modulation

Journal of Structural Biology · doi:10.1016/j.jsb.2013.04.004 ↗ · Activity + Stability - Product Formation

48 measurements ▼

Database ID

UniProt: Q5NLA1 ↗

Sequence Annotation

Explicit - Provided

Protein Source

Recombinant, host bacterium Escherichia coli BL21 (DE3)

Experimental Data (48 measurements)

48 measurements — page 1 of 3

Mutation	Property	Assay	Solvent	Solvent Volume	Aqueous Reference	WT Reference	Measured Value	Units	Solution	pH	Temperature	Substrate(s)	Product(s)	Cofactor(s)	Shaking	Comments
WT	Activity + Stability - Product Formation	Product yield after incubaton (4 hours at 30°C) in the presence of organic solvent, measured by HPLC	Acetone	10%	74.5	—	96.4	%	50 mM Tris-HCl	7.5	30°C	4 mM Ketoisophorone	Levodione	4 mM NADH	180 rpm	Classical aqueous control (in %)
WT	Activity + Stability - Product Formation	Product yield after incubaton (4 hours at 30°C) in the presence of organic solvent, measured by HPLC	Acetone	5%	74.5	—	80.0	%	50 mM Tris-HCl	7.5	30°C	4 mM Ketoisophorone	Levodione	4 mM NADH	180 rpm	Classical aqueous control (in %)
WT	Activity + Stability - Product Formation	Product yield after incubaton (4 hours at 30°C) in the presence of organic solvent, measured by HPLC	Tetrahydrofuran (THF)	30%	74.5	—	1.8	%	50 mM Tris-HCl	7.5	30°C	4 mM Ketoisophorone	Levodione	4 mM NADH	180 rpm	Classical aqueous control (in %)
WT	Activity + Stability - Product Formation	Product yield after incubaton (4 hours at 30°C) in the presence of organic solvent, measured by HPLC	Tetrahydrofuran (THF)	20%	74.5	—	17.3	%	50 mM Tris-HCl	7.5	30°C	4 mM Ketoisophorone	Levodione	4 mM NADH	180 rpm	Classical aqueous control (in %)
WT	Activity + Stability - Product Formation	Product yield after incubaton (4 hours at 30°C) in the presence of organic solvent, measured by HPLC	Tetrahydrofuran (THF)	10%	74.5	—	98.2	%	50 mM Tris-HCl	7.5	30°C	4 mM Ketoisophorone	Levodione	4 mM NADH	180 rpm	Classical aqueous control (in %)
WT	Activity + Stability - Product Formation	Product yield after incubaton (4 hours at 30°C) in the presence of organic solvent, measured by HPLC	Tetrahydrofuran (THF)	5%	74.5	—	90.2	%	50 mM Tris-HCl	7.5	30°C	4 mM Ketoisophorone	Levodione	4 mM NADH	180 rpm	Classical aqueous control (in %)
WT	Activity + Stability - Product Formation	Product yield after incubaton (4 hours at 30°C) in the presence of organic solvent, measured by HPLC	Ethyl Acetate	30%	74.5	—	23.8	%	50 mM Tris-HCl	7.5	30°C	4 mM Ketoisophorone	Levodione	4 mM NADH	180 rpm	Classical aqueous control (in %)
WT	Activity + Stability - Product Formation	Product yield after incubaton (4 hours at 30°C) in the presence of organic solvent, measured by HPLC	Ethyl Acetate	20%	74.5	—	36.3	%	50 mM Tris-HCl	7.5	30°C	4 mM Ketoisophorone	Levodione	4 mM NADH	180 rpm	Classical aqueous control (in %)
WT	Activity + Stability - Product Formation	Product yield after incubaton (4 hours at 30°C) in the presence of organic solvent, measured by HPLC	Ethyl Acetate	10%	74.5	—	65.0	%	50 mM Tris-HCl	7.5	30°C	4 mM Ketoisophorone	Levodione	4 mM NADH	180 rpm	Classical aqueous control (in %)
WT	Activity + Stability - Product Formation	Product yield after incubaton (4 hours at 30°C) in the presence of organic solvent, measured by HPLC	Isopropanol	30%	74.5	—	33.8	%	50 mM Tris-HCl	7.5	30°C	4 mM Ketoisophorone	Levodione	4 mM NADH	180 rpm	Classical aqueous control (in %)
WT	Activity + Stability - Product Formation	Product yield after incubaton (4 hours at 30°C) in the presence of organic solvent, measured by HPLC	Isopropanol	20%	74.5	—	60.4	%	50 mM Tris-HCl	7.5	30°C	4 mM Ketoisophorone	Levodione	4 mM NADH	180 rpm	Classical aqueous control (in %)
WT	Activity + Stability - Product Formation	Product yield after incubaton (4 hours at 30°C) in the presence of organic solvent, measured by HPLC	Isopropanol	10%	74.5	—	64.0	%	50 mM Tris-HCl	7.5	30°C	4 mM Ketoisophorone	Levodione	4 mM NADH	180 rpm	Classical aqueous control (in %)
WT	Activity + Stability - Product Formation	Product yield after incubaton (4 hours at 30°C) in the presence of organic solvent, measured by HPLC	Isopropanol	5%	74.5	—	83.6	%	50 mM Tris-HCl	7.5	30°C	4 mM Ketoisophorone	Levodione	4 mM NADH	180 rpm	Classical aqueous control (in %)
WT	Activity + Stability - Product Formation	Product yield after incubaton (4 hours at 30°C) in the presence of organic solvent, measured by HPLC	Acetone	30%	74.5	—	30.8	%	50 mM Tris-HCl	7.5	30°C	4 mM Ketoisophorone	Levodione	4 mM NADH	180 rpm	Classical aqueous control (in %)
WT	Activity + Stability - Product Formation	Product yield after incubaton (4 hours at 30°C) in the presence of organic solvent, measured by HPLC	Acetone	20%	74.5	—	51.3	%	50 mM Tris-HCl	7.5	30°C	4 mM Ketoisophorone	Levodione	4 mM NADH	180 rpm	Classical aqueous control (in %)
WT	Activity + Stability - Product Formation	Product yield after incubaton (4 hours at 30°C) in the presence of organic solvent, measured by HPLC	Ethyl Acetate	5%	74.5	—	79.8	%	50 mM Tris-HCl	7.5	30°C	4 mM Ketoisophorone	Levodione	4 mM NADH	180 rpm	Classical aqueous control (in %)
DEL267-270	Activity + Stability - Product Formation	Product yield after incubaton (4 hours at 30°C) in the presence of organic solvent, measured by HPLC	Acetone	5%	99.6	80.0	83.5	%	50 mM Tris-HCl	7.5	30°C	4 mM Ketoisophorone	Levodione	4 mM NADH	180 rpm	Classical aqueous control (in %)
DEL267-270	Activity + Stability - Product Formation	Product yield after incubaton (4 hours at 30°C) in the presence of organic solvent, measured by HPLC	Acetone	10%	99.6	96.4	98.2	%	50 mM Tris-HCl	7.5	30°C	4 mM Ketoisophorone	Levodione	4 mM NADH	180 rpm	Classical aqueous control (in %)
DEL267-270	Activity + Stability - Product Formation	Product yield after incubaton (4 hours at 30°C) in the presence of organic solvent, measured by HPLC	Acetone	20%	99.6	51.3	92.4	%	50 mM Tris-HCl	7.5	30°C	4 mM Ketoisophorone	Levodione	4 mM NADH	180 rpm	Classical aqueous control (in %)
DEL267-270	Activity + Stability - Product Formation	Product yield after incubaton (4 hours at 30°C) in the presence of organic solvent, measured by HPLC	Acetone	30%	99.6	30.8	70.1	%	50 mM Tris-HCl	7.5	30°C	4 mM Ketoisophorone	Levodione	4 mM NADH	180 rpm	Classical aqueous control (in %)

Activity + Stability - Product Formation

Product yield after incubaton (4 hours at 30°C) in the presence of organic solvent, measured by HPLC

Acetone

10%

74.5

—

96.4

50 mM Tris-HCl

7.5

30°C

4 mM Ketoisophorone

Levodione

4 mM NADH

180 rpm

Classical aqueous control (in %)

Activity + Stability - Product Formation

Product yield after incubaton (4 hours at 30°C) in the presence of organic solvent, measured by HPLC

Acetone

74.5

—

80.0

50 mM Tris-HCl

7.5

30°C

4 mM Ketoisophorone

Levodione

4 mM NADH

180 rpm

Classical aqueous control (in %)

Activity + Stability - Product Formation

Product yield after incubaton (4 hours at 30°C) in the presence of organic solvent, measured by HPLC

Tetrahydrofuran (THF)

30%

74.5

—

1.8

50 mM Tris-HCl

7.5

30°C

4 mM Ketoisophorone

Levodione

4 mM NADH

180 rpm

Classical aqueous control (in %)

Activity + Stability - Product Formation

Product yield after incubaton (4 hours at 30°C) in the presence of organic solvent, measured by HPLC

Tetrahydrofuran (THF)

20%

74.5

—

17.3

50 mM Tris-HCl

7.5

30°C

4 mM Ketoisophorone

Levodione

4 mM NADH

180 rpm

Classical aqueous control (in %)

Activity + Stability - Product Formation

Product yield after incubaton (4 hours at 30°C) in the presence of organic solvent, measured by HPLC

Tetrahydrofuran (THF)

10%

74.5

—

98.2

50 mM Tris-HCl

7.5

30°C

4 mM Ketoisophorone

Levodione

4 mM NADH

180 rpm

Classical aqueous control (in %)

Activity + Stability - Product Formation

Product yield after incubaton (4 hours at 30°C) in the presence of organic solvent, measured by HPLC

Tetrahydrofuran (THF)

74.5

—

90.2

50 mM Tris-HCl

7.5

30°C

4 mM Ketoisophorone

Levodione

4 mM NADH

180 rpm

Classical aqueous control (in %)

Activity + Stability - Product Formation

Product yield after incubaton (4 hours at 30°C) in the presence of organic solvent, measured by HPLC

Ethyl Acetate

30%

74.5

—

23.8

50 mM Tris-HCl

7.5

30°C

4 mM Ketoisophorone

Levodione

4 mM NADH

180 rpm

Classical aqueous control (in %)

Activity + Stability - Product Formation

Product yield after incubaton (4 hours at 30°C) in the presence of organic solvent, measured by HPLC

Ethyl Acetate

20%

74.5

—

36.3

50 mM Tris-HCl

7.5

30°C

4 mM Ketoisophorone

Levodione

4 mM NADH

180 rpm

Classical aqueous control (in %)

Activity + Stability - Product Formation

Product yield after incubaton (4 hours at 30°C) in the presence of organic solvent, measured by HPLC

Ethyl Acetate

10%

74.5

—

65.0

50 mM Tris-HCl

7.5

30°C

4 mM Ketoisophorone

Levodione

4 mM NADH

180 rpm

Classical aqueous control (in %)

Activity + Stability - Product Formation

Product yield after incubaton (4 hours at 30°C) in the presence of organic solvent, measured by HPLC

Isopropanol

30%

74.5

—

33.8

50 mM Tris-HCl

7.5

30°C

4 mM Ketoisophorone

Levodione

4 mM NADH

180 rpm

Classical aqueous control (in %)

Activity + Stability - Product Formation

Product yield after incubaton (4 hours at 30°C) in the presence of organic solvent, measured by HPLC

Isopropanol

20%

74.5

—

60.4

50 mM Tris-HCl

7.5

30°C

4 mM Ketoisophorone

Levodione

4 mM NADH

180 rpm

Classical aqueous control (in %)

Activity + Stability - Product Formation

Product yield after incubaton (4 hours at 30°C) in the presence of organic solvent, measured by HPLC

Isopropanol

10%

74.5

—

64.0

50 mM Tris-HCl

7.5

30°C

4 mM Ketoisophorone

Levodione

4 mM NADH

180 rpm

Classical aqueous control (in %)

Activity + Stability - Product Formation

Product yield after incubaton (4 hours at 30°C) in the presence of organic solvent, measured by HPLC

Isopropanol

74.5

—

83.6

50 mM Tris-HCl

7.5

30°C

4 mM Ketoisophorone

Levodione

4 mM NADH

180 rpm

Classical aqueous control (in %)

Activity + Stability - Product Formation

Product yield after incubaton (4 hours at 30°C) in the presence of organic solvent, measured by HPLC

Acetone

30%

74.5

—

30.8

50 mM Tris-HCl

7.5

30°C

4 mM Ketoisophorone

Levodione

4 mM NADH

180 rpm

Classical aqueous control (in %)

Activity + Stability - Product Formation

Product yield after incubaton (4 hours at 30°C) in the presence of organic solvent, measured by HPLC

Acetone

20%

74.5

—

51.3

50 mM Tris-HCl

7.5

30°C

4 mM Ketoisophorone

Levodione

4 mM NADH

180 rpm

Classical aqueous control (in %)

Activity + Stability - Product Formation

Product yield after incubaton (4 hours at 30°C) in the presence of organic solvent, measured by HPLC

Ethyl Acetate

74.5

—

79.8

50 mM Tris-HCl

7.5

30°C

4 mM Ketoisophorone

Levodione

4 mM NADH

180 rpm

Classical aqueous control (in %)

DEL267-270

Activity + Stability - Product Formation

Product yield after incubaton (4 hours at 30°C) in the presence of organic solvent, measured by HPLC

Acetone

99.6

80.0

83.5

50 mM Tris-HCl

7.5

30°C

4 mM Ketoisophorone

Levodione

4 mM NADH

180 rpm

Classical aqueous control (in %)

DEL267-270

Activity + Stability - Product Formation

Product yield after incubaton (4 hours at 30°C) in the presence of organic solvent, measured by HPLC

Acetone

10%

99.6

96.4

98.2

50 mM Tris-HCl

7.5

30°C

4 mM Ketoisophorone

Levodione

4 mM NADH

180 rpm

Classical aqueous control (in %)

DEL267-270

Activity + Stability - Product Formation

Product yield after incubaton (4 hours at 30°C) in the presence of organic solvent, measured by HPLC

Acetone

20%

99.6

51.3

92.4

50 mM Tris-HCl

7.5

30°C

4 mM Ketoisophorone

Levodione

4 mM NADH

180 rpm

Classical aqueous control (in %)

DEL267-270

Activity + Stability - Product Formation

Product yield after incubaton (4 hours at 30°C) in the presence of organic solvent, measured by HPLC

Acetone

30%

99.6

30.8

70.1

50 mM Tris-HCl

7.5

30°C

4 mM Ketoisophorone

Levodione

4 mM NADH

180 rpm

Classical aqueous control (in %)

DEL267-270

Activity + Stability - Product Formation

Product yield after incubaton (4 hours at 30°C) in the presence of organic solvent, measured by HPLC

Isopropanol

99.6

83.6

81.3

50 mM Tris-HCl

7.5

30°C

4 mM Ketoisophorone

Levodione

4 mM NADH

180 rpm

Classical aqueous control (in %)

DEL267-270

Activity + Stability - Product Formation

Product yield after incubaton (4 hours at 30°C) in the presence of organic solvent, measured by HPLC

Isopropanol

10%

99.6

64.0

85.3

50 mM Tris-HCl

7.5

30°C

4 mM Ketoisophorone

Levodione

4 mM NADH

180 rpm

Classical aqueous control (in %)

DEL267-270

Activity + Stability - Product Formation

Product yield after incubaton (4 hours at 30°C) in the presence of organic solvent, measured by HPLC

Isopropanol

20%

99.6

60.4

79.6

50 mM Tris-HCl

7.5

30°C

4 mM Ketoisophorone

Levodione

4 mM NADH

180 rpm

Classical aqueous control (in %)

DEL267-270

Activity + Stability - Product Formation

Product yield after incubaton (4 hours at 30°C) in the presence of organic solvent, measured by HPLC

Isopropanol

30%

99.6

33.8

68.0

50 mM Tris-HCl

7.5

30°C

4 mM Ketoisophorone

Levodione

4 mM NADH

180 rpm

Classical aqueous control (in %)

DEL267-270

Activity + Stability - Product Formation

Product yield after incubaton (4 hours at 30°C) in the presence of organic solvent, measured by HPLC

Ethyl Acetate

99.6

79.8

80.3

50 mM Tris-HCl

7.5

30°C

4 mM Ketoisophorone

Levodione

4 mM NADH

180 rpm

Classical aqueous control (in %)

DEL267-270

Activity + Stability - Product Formation

Product yield after incubaton (4 hours at 30°C) in the presence of organic solvent, measured by HPLC

Ethyl Acetate

10%

99.6

65.0

87.4

50 mM Tris-HCl

7.5

30°C

4 mM Ketoisophorone

Levodione

4 mM NADH

180 rpm

Classical aqueous control (in %)

DEL267-270

Activity + Stability - Product Formation

Product yield after incubaton (4 hours at 30°C) in the presence of organic solvent, measured by HPLC

Ethyl Acetate

20%

99.6

36.3

74.4

50 mM Tris-HCl

7.5

30°C

4 mM Ketoisophorone

Levodione

4 mM NADH

180 rpm

Classical aqueous control (in %)

DEL267-270

Activity + Stability - Product Formation

Product yield after incubaton (4 hours at 30°C) in the presence of organic solvent, measured by HPLC

Ethyl Acetate

30%

99.6

23.8

57.4

50 mM Tris-HCl

7.5

30°C

4 mM Ketoisophorone

Levodione

4 mM NADH

180 rpm

Classical aqueous control (in %)

DEL267-270

Activity + Stability - Product Formation

Product yield after incubaton (4 hours at 30°C) in the presence of organic solvent, measured by HPLC

Tetrahydrofuran (THF)

99.6

90.2

98.2

50 mM Tris-HCl

7.5

30°C

4 mM Ketoisophorone

Levodione

4 mM NADH

180 rpm

Classical aqueous control (in %)

DEL267-270

Activity + Stability - Product Formation

Product yield after incubaton (4 hours at 30°C) in the presence of organic solvent, measured by HPLC

Tetrahydrofuran (THF)

10%

99.6

98.2

65.3

50 mM Tris-HCl

7.5

30°C

4 mM Ketoisophorone

Levodione

4 mM NADH

180 rpm

Classical aqueous control (in %)

DEL267-270

Activity + Stability - Product Formation

Product yield after incubaton (4 hours at 30°C) in the presence of organic solvent, measured by HPLC

Tetrahydrofuran (THF)

20%

99.6

17.3

41.3

50 mM Tris-HCl

7.5

30°C

4 mM Ketoisophorone

Levodione

4 mM NADH

180 rpm

Classical aqueous control (in %)

DEL267-270

Activity + Stability - Product Formation

Product yield after incubaton (4 hours at 30°C) in the presence of organic solvent, measured by HPLC

Tetrahydrofuran (THF)

30%

99.6

1.8

12.0

50 mM Tris-HCl

7.5

30°C

4 mM Ketoisophorone

Levodione

4 mM NADH

180 rpm

Classical aqueous control (in %)

DEL230-236

Activity + Stability - Product Formation

Product yield after incubaton (4 hours at 30°C) in the presence of organic solvent, measured by HPLC

Tetrahydrofuran (THF)

30%

97.3

1.8

1.3

50 mM Tris-HCl

7.5

30°C

4 mM Ketoisophorone

Levodione

4 mM NADH

180 rpm

Classical aqueous control (in %)

DEL230-236

Activity + Stability - Product Formation

Product yield after incubaton (4 hours at 30°C) in the presence of organic solvent, measured by HPLC

Tetrahydrofuran (THF)

20%

97.3

17.3

1.3

50 mM Tris-HCl

7.5

30°C

4 mM Ketoisophorone

Levodione

4 mM NADH

180 rpm

Classical aqueous control (in %)

DEL230-236

Activity + Stability - Product Formation

Product yield after incubaton (4 hours at 30°C) in the presence of organic solvent, measured by HPLC

Tetrahydrofuran (THF)

10%

97.3

98.2

11.1

50 mM Tris-HCl

7.5

30°C

4 mM Ketoisophorone

Levodione

4 mM NADH

180 rpm

Classical aqueous control (in %)

DEL230-236

Activity + Stability - Product Formation

Product yield after incubaton (4 hours at 30°C) in the presence of organic solvent, measured by HPLC

Tetrahydrofuran (THF)

97.3

90.2

54.7

50 mM Tris-HCl

7.5

30°C

4 mM Ketoisophorone

Levodione

4 mM NADH

180 rpm

Classical aqueous control (in %)

DEL230-236

Activity + Stability - Product Formation

Product yield after incubaton (4 hours at 30°C) in the presence of organic solvent, measured by HPLC

Ethyl Acetate

30%

97.3

23.8

16.1

50 mM Tris-HCl

7.5

30°C

4 mM Ketoisophorone

Levodione

4 mM NADH

180 rpm

Classical aqueous control (in %)

DEL230-236

Activity + Stability - Product Formation

Product yield after incubaton (4 hours at 30°C) in the presence of organic solvent, measured by HPLC

Ethyl Acetate

20%

97.3

36.3

4.9

50 mM Tris-HCl

7.5

30°C

4 mM Ketoisophorone

Levodione

4 mM NADH

180 rpm

Classical aqueous control (in %)

DEL230-236

Activity + Stability - Product Formation

Product yield after incubaton (4 hours at 30°C) in the presence of organic solvent, measured by HPLC

Ethyl Acetate

10%

97.3

65.0

12.1

50 mM Tris-HCl

7.5

30°C

4 mM Ketoisophorone

Levodione

4 mM NADH

180 rpm

Classical aqueous control (in %)

DEL230-236

Activity + Stability - Product Formation

Product yield after incubaton (4 hours at 30°C) in the presence of organic solvent, measured by HPLC

Ethyl Acetate

97.3

79.8

35.9

50 mM Tris-HCl

7.5

30°C

4 mM Ketoisophorone

Levodione

4 mM NADH

180 rpm

Classical aqueous control (in %)

DEL230-236

Activity + Stability - Product Formation

Product yield after incubaton (4 hours at 30°C) in the presence of organic solvent, measured by HPLC

Isopropanol

30%

97.3

33.8

15.6

50 mM Tris-HCl

7.5

30°C

4 mM Ketoisophorone

Levodione

4 mM NADH

180 rpm

Classical aqueous control (in %)

DEL230-236

Activity + Stability - Product Formation

Product yield after incubaton (4 hours at 30°C) in the presence of organic solvent, measured by HPLC

Isopropanol

20%

97.3

60.4

48.4

50 mM Tris-HCl

7.5

30°C

4 mM Ketoisophorone

Levodione

4 mM NADH

180 rpm

Classical aqueous control (in %)

DEL230-236

Activity + Stability - Product Formation

Product yield after incubaton (4 hours at 30°C) in the presence of organic solvent, measured by HPLC

Isopropanol

10%

97.3

64.0

52.4

50 mM Tris-HCl

7.5

30°C

4 mM Ketoisophorone

Levodione

4 mM NADH

180 rpm

Classical aqueous control (in %)

DEL230-236

Activity + Stability - Product Formation

Product yield after incubaton (4 hours at 30°C) in the presence of organic solvent, measured by HPLC

Isopropanol

97.3

83.6

69.3

50 mM Tris-HCl

7.5

30°C

4 mM Ketoisophorone

Levodione

4 mM NADH

180 rpm

Classical aqueous control (in %)

DEL230-236

Activity + Stability - Product Formation

Product yield after incubaton (4 hours at 30°C) in the presence of organic solvent, measured by HPLC

Acetone

30%

97.3

30.8

3.1

50 mM Tris-HCl

7.5

30°C

4 mM Ketoisophorone

Levodione

4 mM NADH

180 rpm

Classical aqueous control (in %)

DEL230-236

Activity + Stability - Product Formation

Product yield after incubaton (4 hours at 30°C) in the presence of organic solvent, measured by HPLC

Acetone

20%

97.3

51.3

10.3

50 mM Tris-HCl

7.5

30°C

4 mM Ketoisophorone

Levodione

4 mM NADH

180 rpm

Classical aqueous control (in %)

DEL230-236

Activity + Stability - Product Formation

Product yield after incubaton (4 hours at 30°C) in the presence of organic solvent, measured by HPLC

Acetone

10%

97.3

96.4

51.8

50 mM Tris-HCl

7.5

30°C

4 mM Ketoisophorone

Levodione

4 mM NADH

180 rpm

Classical aqueous control (in %)

DEL230-236

Activity + Stability - Product Formation

Product yield after incubaton (4 hours at 30°C) in the presence of organic solvent, measured by HPLC

Acetone

97.3

80.0

66.5

50 mM Tris-HCl

7.5

30°C

4 mM Ketoisophorone

Levodione

4 mM NADH

180 rpm

Classical aqueous control (in %)

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Mutations in this dataset (3) — Sabrina Reich et al. (2014)

WT DEL230-236 DEL267-270

Visualization : Activity + Stability — Product Formation

Sabrina Reich et al. (2014)

One bar per measurement. Colour = solvent, shade = solvent volume. — — — Reference value. Hover for details.

Mutation Effect

Mutation impact on enzyme stability and function in the presence of organic solvent: comparison of wild-type and mutant values in identical conditions.

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: Q5NLA1 ↗

Sequence Annotation

Explicit - Provided UniProt Accession Number

Protein Source

Recombinant, host bacterium Escherichia coli BL21 (DE3)

Experimental Data (150 measurements)

150 measurements — page 5 of 8

Property	Assay	Solvent	Solvent Volume	Aqueous Reference	Measured Value	Units	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	Methanol	30%	100.0	34.8	%	50 mM sodium phosphate	7.4	25	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%	100.0	15.2	%	50 mM sodium phosphate	7.4	25	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%	100.0	8.7	%	50 mM sodium phosphate	7.4	25	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%	100.0	2.2	%	50 mM sodium phosphate	7.4	25	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%	100.0	93.5	%	50 mM sodium phosphate	7.4	30	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%	100.0	79.0	%	50 mM sodium phosphate	7.4	30	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%	100.0	66.1	%	50 mM sodium phosphate	7.4	30	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%	100.0	50.0	%	50 mM sodium phosphate	7.4	30	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%	100.0	38.7	%	50 mM sodium phosphate	7.4	30	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%	100.0	21.0	%	50 mM sodium phosphate	7.4	30	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%	100.0	6.5	%	50 mM sodium phosphate	7.4	30	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%	100.0	4.8	%	50 mM sodium phosphate	7.4	30	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%	100.0	8.1	%	50 mM sodium phosphate	7.4	30	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%	100.0	96.7	%	50 mM sodium phosphate	7.4	35	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%	100.0	78.7	%	50 mM sodium phosphate	7.4	35	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%	100.0	67.2	%	50 mM sodium phosphate	7.4	35	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%	100.0	45.9	%	50 mM sodium phosphate	7.4	35	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%	100.0	24.6	%	50 mM sodium phosphate	7.4	35	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%	100.0	3.3	%	50 mM sodium phosphate	7.4	35	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%	100.0	1.6	%	50 mM sodium phosphate	7.4	35	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	Dimethyl Sulfoxide (DMSO)	5%	100.0	83.0	%	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%	100.0	63.8	%	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%	100.0	48.9	%	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%	100.0	31.9	%	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%	100.0	21.3	%	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%	100.0	10.6	%	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%	100.0	83.0	%	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%	100.0	76.6	%	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%	100.0	59.6	%	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%	100.0	44.7	%	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%	100.0	31.9	%	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%	100.0	19.1	%	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%	100.0	93.5	%	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%	100.0	80.4	%	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%	100.0	71.7	%	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%	100.0	60.9	%	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%	100.0	47.8	%	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%	100.0	39.1	%	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%	100.0	84.1	%	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%	100.0	77.3	%	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%	100.0	56.8	%	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%	100.0	38.6	%	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%	100.0	31.8	%	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%	100.0	29.5	%	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%	100.0	68.9	%	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%	100.0	44.4	%	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%	100.0	31.1	%	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%	100.0	22.2	%	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%	100.0	15.6	%	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%	100.0	13.3	%	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%	100.0	97.8	%	50 mM sodium phosphate	7.4	25	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%	100.0	82.2	%	50 mM sodium phosphate	7.4	25	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%	100.0	64.4	%	50 mM sodium phosphate	7.4	25	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%	100.0	44.4	%	50 mM sodium phosphate	7.4	25	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%	100.0	24.4	%	50 mM sodium phosphate	7.4	25	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%	100.0	20.0	%	50 mM sodium phosphate	7.4	25	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%	100.0	100.0	%	50 mM sodium phosphate	7.4	25	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%	100.0	4.4	%	50 mM sodium phosphate	7.4	25	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%	100.0	0.0	%	50 mM sodium phosphate	7.4	25	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%	100.0	91.4	%	50 mM sodium phosphate	7.4	30	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%	100.0	70.7	%	50 mM sodium phosphate	7.4	30	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%	100.0	53.4	%	50 mM sodium phosphate	7.4	30	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%	100.0	36.2	%	50 mM sodium phosphate	7.4	30	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%	100.0	20.7	%	50 mM sodium phosphate	7.4	30	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%	100.0	8.6	%	50 mM sodium phosphate	7.4	30	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%	100.0	5.2	%	50 mM sodium phosphate	7.4	30	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%	100.0	0.0	%	50 mM sodium phosphate	7.4	30	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%	100.0	0.0	%	50 mM sodium phosphate	7.4	30	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%	100.0	87.7	%	50 mM sodium phosphate	7.4	35	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%	100.0	73.8	%	50 mM sodium phosphate	7.4	35	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%	100.0	46.2	%	50 mM sodium phosphate	7.4	35	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%	100.0	18.5	%	50 mM sodium phosphate	7.4	35	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%	100.0	9.2	%	50 mM sodium phosphate	7.4	35	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%	100.0	3.1	%	50 mM sodium phosphate	7.4	35	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%	100.0	4.6	%	50 mM sodium phosphate	7.4	35	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%	100.0	7.7	%	50 mM sodium phosphate	7.4	35	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%	100.0	13.8	%	50 mM sodium phosphate	7.4	35	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%	100.0	80.5	%	50 mM sodium phosphate	7.4	40	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%	100.0	46.3	%	50 mM sodium phosphate	7.4	40	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%	100.0	17.1	%	50 mM sodium phosphate	7.4	40	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%	100.0	4.9	%	50 mM sodium phosphate	7.4	40	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%	100.0	4.9	%	50 mM sodium phosphate	7.4	40	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%	100.0	4.9	%	50 mM sodium phosphate	7.4	40	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%	100.0	7.3	%	50 mM sodium phosphate	7.4	40	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%	100.0	4.9	%	50 mM sodium phosphate	7.4	40	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%	100.0	12.2	%	50 mM sodium phosphate	7.4	40	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%	100.0	33.3	%	50 mM sodium phosphate	7.4	45	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%	100.0	116.7	%	50 mM sodium phosphate	7.4	45	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%	100.0	25.0	%	50 mM sodium phosphate	7.4	45	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%	100.0	8.3	%	50 mM sodium phosphate	7.4	45	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%	100.0	50.0	%	50 mM sodium phosphate	7.4	45	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%	100.0	16.7	%	50 mM sodium phosphate	7.4	45	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%	100.0	41.7	%	50 mM sodium phosphate	7.4	45	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%	100.0	66.7	%	50 mM sodium phosphate	7.4	45	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%	100.0	25.0	%	50 mM sodium phosphate	7.4	45	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%	100.0	102.2	%	50 mM sodium phosphate	7.4	25	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%	100.0	97.8	%	50 mM sodium phosphate	7.4	25	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%	100.0	82.6	%	50 mM sodium phosphate	7.4	25	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%	100.0	69.6	%	50 mM sodium phosphate	7.4	25	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%	100.0	47.8	%	50 mM sodium phosphate	7.4	25	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%	100.0	34.8	%	50 mM sodium phosphate	7.4	25	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%	100.0	15.2	%	50 mM sodium phosphate	7.4	25	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%	100.0	8.7	%	50 mM sodium phosphate	7.4	25	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%	100.0	2.2	%	50 mM sodium phosphate	7.4	25	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%	100.0	93.5	%	50 mM sodium phosphate	7.4	30	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%	100.0	79.0	%	50 mM sodium phosphate	7.4	30	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%	100.0	66.1	%	50 mM sodium phosphate	7.4	30	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%	100.0	50.0	%	50 mM sodium phosphate	7.4	30	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%	100.0	38.7	%	50 mM sodium phosphate	7.4	30	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%	100.0	21.0	%	50 mM sodium phosphate	7.4	30	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%	100.0	6.5	%	50 mM sodium phosphate	7.4	30	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%	100.0	4.8	%	50 mM sodium phosphate	7.4	30	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%	100.0	8.1	%	50 mM sodium phosphate	7.4	30	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%	100.0	96.7	%	50 mM sodium phosphate	7.4	35	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%	100.0	78.7	%	50 mM sodium phosphate	7.4	35	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%	100.0	67.2	%	50 mM sodium phosphate	7.4	35	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%	100.0	45.9	%	50 mM sodium phosphate	7.4	35	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%	100.0	24.6	%	50 mM sodium phosphate	7.4	35	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%	100.0	3.3	%	50 mM sodium phosphate	7.4	35	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%	100.0	1.6	%	50 mM sodium phosphate	7.4	35	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%	100.0	0.0	%	50 mM sodium phosphate	7.4	35	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%	100.0	3.3	%	50 mM sodium phosphate	7.4	35	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%	100.0	76.7	%	50 mM sodium phosphate	7.4	40	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%	100.0	53.5	%	50 mM sodium phosphate	7.4	40	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%	100.0	32.6	%	50 mM sodium phosphate	7.4	40	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%	100.0	7.0	%	50 mM sodium phosphate	7.4	40	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%	100.0	0.0	%	50 mM sodium phosphate	7.4	40	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%	100.0	2.3	%	50 mM sodium phosphate	7.4	40	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%	100.0	2.3	%	50 mM sodium phosphate	7.4	40	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%	100.0	2.3	%	50 mM sodium phosphate	7.4	40	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%	100.0	7.0	%	50 mM sodium phosphate	7.4	40	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%	100.0	100.0	%	50 mM sodium phosphate	7.4	45	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%	100.0	0.0	%	50 mM sodium phosphate	7.4	45	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%	100.0	100.0	%	50 mM sodium phosphate	7.4	45	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%	100.0	0.0	%	50 mM sodium phosphate	7.4	45	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%	100.0	0.0	%	50 mM sodium phosphate	7.4	45	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%	100.0	25.0	%	50 mM sodium phosphate	7.4	45	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%	100.0	100.0	%	50 mM sodium phosphate	7.4	45	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%	100.0	100.0	%	50 mM sodium phosphate	7.4	45	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%	100.0	75.0	%	50 mM sodium phosphate	7.4	45	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%	50.33	49.0	°C	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%	50.33	47.0	°C	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%	50.33	45.33	°C	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%	50.33	43.33	°C	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%	50.33	42.67	°C	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%	50.33	41.33	°C	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%	50.33	47.67	°C	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%	50.33	43.33	°C	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%	50.33	40.33	°C	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%	50.33	38.0	°C	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%	50.33	35.67	°C	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%	50.33	34.33	°C	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%	50.33	49.0	°C	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%	50.33	47.33	°C	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%	50.33	45.67	°C	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%	50.33	42.33	°C	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%	50.33	40.0	°C	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%	50.33	39.0	°C	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%	50.33	47.33	°C	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%	50.33	43.33	°C	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%	50.33	39.67	°C	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%	50.33	35.67	°C	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%	50.33	33.0	°C	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%	50.33	30.0	°C	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%	50.33	44.33	°C	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%	50.33	37.33	°C	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%	50.33	33.33	°C	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%	50.33	31.33	°C	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%	50.33	30.33	°C	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%	50.33	30.0	°C	50 mM sodium phosphate	7.4	—	—	—	—	—	Classical aqueous control (in °C)

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Visualization : Activity — Classical

Frieda A. Sorgenfrei et al. (2024)

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

Visualization : Stability — T_m

Frieda A. Sorgenfrei et al. (2024)

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

Structure

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Ene-reductase NCR from Zymomonas mobilis

Enzyme Description

Sequence

Experimental Data (48 measurements)

Visualization : Activity + Stability — Product Formation

Mutation Effect

Experimental Data (150 measurements)

Visualization : Activity — Classical

Visualization : Stability — Tm

Structure

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Visualization : Stability — T_m