3-hydroxy-3-methylglutaryl coenzyme A reductase from Sulfolobus solfataricus

Enzyme Description

Species

Sulfolobus solfataricus ↗

Extremophile

Yes "thermoacidophilic" organism

EC Number

1.1.1.34 ↗

Sequence

Length: 409 amino acids

MKIDEVVEKLVKGEISFHEVDNLLEANAAMVARRLALEKIVGVGLPSIGSTVIDYSEIKNKNAENVIGAIQIPLGIVGPIRVNGDYAKGDFYVPMATTEGALIASVNRGIKAVTLSGGVRAKVLKDEMTRAPVFKFDSIEQIPNFLKFIEENLEKIRNIANSTSHHGKLKSITPFVLGNNVWLRFSFETGDAMGMNMVTIAVEKVCEFIEENFPSADCLAVSGNMCSDKKQTNVNSLFGRGKTVVAEALIKKDVIRNILHSNAQLIHDINLRKNWLGTARAGSLSQFNAHFANIVTAIFIATGQDVAQIVESSSGYTWTEVRGEDLYISVTLPSLEVGTVGGGTRLPTQKEALSIMGVYGSGNPPGSNAKKLAEIIASTVLSGELNLLAALSNKELGKAHAKLGRAMKV

Michael Dirkmann et al. (2017) — A Multiperspective Approach to Solvent Regulation of Enzymatic Activity: HMG-CoA Reductase

ChemBioChem · doi:10.1002/cbic.201700596 ↗ · Activity - Classical

79 measurements ▼

Database ID

UniProt: O08424 ↗

Sequence Annotation

Explicit - Provided GenBank Accession Number

Protein Source

Recombinant, host bacterium Escherichia coli Rosetta‐gamiTM 2(DE3)

Experimental Data (79 measurements)

79 measurements — page 3 of 4

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 (DTNB and released coenzyme A absorbance measurement, 412 nm) in the presence of organic solvent	Tetrahydrofuran (THF)	5 % v/v	100	109	%	Digital	Continuous	100 mM KOAc/100 mM MES buffer, 0.25 mM 5.5-dithio-bis-(2-nitrobenzoic acid)	5.5	45°C	0.25 mM Hydroxymethylglutaroyl coenzyme A	Coenzyme A , Mevalonate	0.5 mM NADPH	—	Classical aqueous control (in %) \| Racemic substrate
Activity - Classical	Activity measured by absorbance spectrophotometry (DTNB and released coenzyme A absorbance measurement, 412 nm) in the presence of organic solvent	Dimethyl Sulfoxide (DMSO)	5 % v/v	100	89	%	Digital	Continuous	100 mM KOAc/100 mM MES buffer, 0.25 mM 5.5-dithio-bis-(2-nitrobenzoic acid)	5.5	45°C	0.25 mM Hydroxymethylglutaroyl coenzyme A	Coenzyme A , Mevalonate	0.5 mM NADPH	—	Classical aqueous control (in %) \| Racemic substrate
Activity - Classical	Activity measured by absorbance spectrophotometry (DTNB and released coenzyme A absorbance measurement, 412 nm) in the presence of organic solvent	Dimethyl Sulfoxide (DMSO)	2.5 % v/v	100	105	%	Digital	Continuous	100 mM KOAc/100 mM MES buffer, 0.25 mM 5.5-dithio-bis-(2-nitrobenzoic acid)	5.5	45°C	0.25 mM Hydroxymethylglutaroyl coenzyme A	Coenzyme A , Mevalonate	0.5 mM NADPH	—	Classical aqueous control (in %) \| Racemic substrate
Activity - Classical	Activity measured by absorbance spectrophotometry (DTNB and released coenzyme A absorbance measurement, 412 nm) in the presence of organic solvent	Dimethyl Sulfoxide (DMSO)	1 % v/v	100	110	%	Digital	Continuous	100 mM KOAc/100 mM MES buffer, 0.25 mM 5.5-dithio-bis-(2-nitrobenzoic acid)	5.5	45°C	0.25 mM Hydroxymethylglutaroyl coenzyme A	Coenzyme A , Mevalonate	0.5 mM NADPH	—	Classical aqueous control (in %) \| Racemic substrate
Activity - Classical	Activity measured by absorbance spectrophotometry (DTNB and released coenzyme A absorbance measurement, 412 nm) in the presence of organic solvent	Tetrahydrofuran (THF)	60 % v/v	100	44	%	Digital	Continuous	100 mM KOAc/100 mM MES buffer, 0.25 mM 5.5-dithio-bis-(2-nitrobenzoic acid)	5.5	45°C	0.25 mM Hydroxymethylglutaroyl coenzyme A	Coenzyme A , Mevalonate	0.5 mM NADPH	—	Classical aqueous control (in %) \| Racemic substrate
Activity - Classical	Activity measured by absorbance spectrophotometry (DTNB and released coenzyme A absorbance measurement, 412 nm) in the presence of organic solvent	Tetrahydrofuran (THF)	50 % v/v	100	48	%	Digital	Continuous	100 mM KOAc/100 mM MES buffer, 0.25 mM 5.5-dithio-bis-(2-nitrobenzoic acid)	5.5	45°C	0.25 mM Hydroxymethylglutaroyl coenzyme A	Coenzyme A , Mevalonate	0.5 mM NADPH	—	Classical aqueous control (in %) \| Racemic substrate
Activity - Classical	Activity measured by absorbance spectrophotometry (DTNB and released coenzyme A absorbance measurement, 412 nm) in the presence of organic solvent	Tetrahydrofuran (THF)	40 % v/v	100	68	%	Digital	Continuous	100 mM KOAc/100 mM MES buffer, 0.25 mM 5.5-dithio-bis-(2-nitrobenzoic acid)	5.5	45°C	0.25 mM Hydroxymethylglutaroyl coenzyme A	Coenzyme A , Mevalonate	0.5 mM NADPH	—	Classical aqueous control (in %) \| Racemic substrate
Activity - Classical	Activity measured by absorbance spectrophotometry (DTNB and released coenzyme A absorbance measurement, 412 nm) in the presence of organic solvent	Tetrahydrofuran (THF)	30 % v/v	100	86	%	Digital	Continuous	100 mM KOAc/100 mM MES buffer, 0.25 mM 5.5-dithio-bis-(2-nitrobenzoic acid)	5.5	45°C	0.25 mM Hydroxymethylglutaroyl coenzyme A	Coenzyme A , Mevalonate	0.5 mM NADPH	—	Classical aqueous control (in %) \| Racemic substrate
Activity - Classical	Activity measured by absorbance spectrophotometry (DTNB and released coenzyme A absorbance measurement, 412 nm) in the presence of organic solvent	Tetrahydrofuran (THF)	20 % v/v	100	98	%	Digital	Continuous	100 mM KOAc/100 mM MES buffer, 0.25 mM 5.5-dithio-bis-(2-nitrobenzoic acid)	5.5	45°C	0.25 mM Hydroxymethylglutaroyl coenzyme A	Coenzyme A , Mevalonate	0.5 mM NADPH	—	Classical aqueous control (in %) \| Racemic substrate
Activity - Classical	Activity measured by absorbance spectrophotometry (DTNB and released coenzyme A absorbance measurement, 412 nm) in the presence of organic solvent	Tetrahydrofuran (THF)	10 % v/v	100	107	%	Digital	Continuous	100 mM KOAc/100 mM MES buffer, 0.25 mM 5.5-dithio-bis-(2-nitrobenzoic acid)	5.5	45°C	0.25 mM Hydroxymethylglutaroyl coenzyme A	Coenzyme A , Mevalonate	0.5 mM NADPH	—	Classical aqueous control (in %) \| Racemic substrate
Activity - Classical	Activity measured by absorbance spectrophotometry (DTNB and released coenzyme A absorbance measurement, 412 nm) in the presence of organic solvent	Dimethyl Sulfoxide (DMSO)	10 % v/v	100	72	%	Digital	Continuous	100 mM KOAc/100 mM MES buffer, 0.25 mM 5.5-dithio-bis-(2-nitrobenzoic acid)	5.5	45°C	0.25 mM Hydroxymethylglutaroyl coenzyme A	Coenzyme A , Mevalonate	0.5 mM NADPH	—	Classical aqueous control (in %) \| Racemic substrate
Activity - Classical	Activity measured by absorbance spectrophotometry (DTNB and released coenzyme A absorbance measurement, 412 nm) in the presence of organic solvent	Tetrahydrofuran (THF)	2.5 % v/v	100	106	%	Digital	Continuous	100 mM KOAc/100 mM MES buffer, 0.25 mM 5.5-dithio-bis-(2-nitrobenzoic acid)	5.5	45°C	0.25 mM Hydroxymethylglutaroyl coenzyme A	Coenzyme A , Mevalonate	0.5 mM NADPH	—	Classical aqueous control (in %) \| Racemic substrate
Activity - Classical	Activity measured by absorbance spectrophotometry (DTNB and released coenzyme A absorbance measurement, 412 nm) in the presence of organic solvent	Acetonitrile	60 % v/v	100	35	%	Digital	Continuous	100 mM KOAc/100 mM MES buffer, 0.25 mM 5.5-dithio-bis-(2-nitrobenzoic acid)	5.5	45°C	0.25 mM Hydroxymethylglutaroyl coenzyme A	Coenzyme A , Mevalonate	0.5 mM NADPH	—	Classical aqueous control (in %) \| Racemic substrate
Activity - Classical	Activity measured by absorbance spectrophotometry (DTNB and released coenzyme A absorbance measurement, 412 nm) in the presence of organic solvent	Acetonitrile	50 % v/v	100	51	%	Digital	Continuous	100 mM KOAc/100 mM MES buffer, 0.25 mM 5.5-dithio-bis-(2-nitrobenzoic acid)	5.5	45°C	0.25 mM Hydroxymethylglutaroyl coenzyme A	Coenzyme A , Mevalonate	0.5 mM NADPH	—	Classical aqueous control (in %) \| Racemic substrate
Activity - Classical	Activity measured by absorbance spectrophotometry (DTNB and released coenzyme A absorbance measurement, 412 nm) in the presence of organic solvent	Acetonitrile	40 % v/v	100	52	%	Digital	Continuous	100 mM KOAc/100 mM MES buffer, 0.25 mM 5.5-dithio-bis-(2-nitrobenzoic acid)	5.5	45°C	0.25 mM Hydroxymethylglutaroyl coenzyme A	Coenzyme A , Mevalonate	0.5 mM NADPH	—	Classical aqueous control (in %) \| Racemic substrate
Activity - Classical	Activity measured by absorbance spectrophotometry (DTNB and released coenzyme A absorbance measurement, 412 nm) in the presence of organic solvent	Acetonitrile	30 % v/v	100	77	%	Digital	Continuous	100 mM KOAc/100 mM MES buffer, 0.25 mM 5.5-dithio-bis-(2-nitrobenzoic acid)	5.5	45°C	0.25 mM Hydroxymethylglutaroyl coenzyme A	Coenzyme A , Mevalonate	0.5 mM NADPH	—	Classical aqueous control (in %) \| Racemic substrate
Activity - Classical	Activity measured by absorbance spectrophotometry (DTNB and released coenzyme A absorbance measurement, 412 nm) in the presence of organic solvent	Acetonitrile	20 % v/v	100	101	%	Digital	Continuous	100 mM KOAc/100 mM MES buffer, 0.25 mM 5.5-dithio-bis-(2-nitrobenzoic acid)	5.5	45°C	0.25 mM Hydroxymethylglutaroyl coenzyme A	Coenzyme A , Mevalonate	0.5 mM NADPH	—	Classical aqueous control (in %) \| Racemic substrate
Activity - Classical	Activity measured by absorbance spectrophotometry (DTNB and released coenzyme A absorbance measurement, 412 nm) in the presence of organic solvent	Acetonitrile	15 % v/v	100	106	%	Digital	Continuous	100 mM KOAc/100 mM MES buffer, 0.25 mM 5.5-dithio-bis-(2-nitrobenzoic acid)	5.5	45°C	0.25 mM Hydroxymethylglutaroyl coenzyme A	Coenzyme A , Mevalonate	0.5 mM NADPH	—	Classical aqueous control (in %) \| Racemic substrate
Activity - Classical	Activity measured by absorbance spectrophotometry (DTNB and released coenzyme A absorbance measurement, 412 nm) in the presence of organic solvent	Acetonitrile	10 % v/v	100	117	%	Digital	Continuous	100 mM KOAc/100 mM MES buffer, 0.25 mM 5.5-dithio-bis-(2-nitrobenzoic acid)	5.5	45°C	0.25 mM Hydroxymethylglutaroyl coenzyme A	Coenzyme A , Mevalonate	0.5 mM NADPH	—	Classical aqueous control (in %) \| Racemic substrate
Activity - Classical	Activity measured by absorbance spectrophotometry (DTNB and released coenzyme A absorbance measurement, 412 nm) in the presence of organic solvent	Acetonitrile	5 % v/v	100	111	%	Digital	Continuous	100 mM KOAc/100 mM MES buffer, 0.25 mM 5.5-dithio-bis-(2-nitrobenzoic acid)	5.5	45°C	0.25 mM Hydroxymethylglutaroyl coenzyme A	Coenzyme A , Mevalonate	0.5 mM NADPH	—	Classical aqueous control (in %) \| Racemic substrate

Activity - Classical

Activity measured by absorbance spectrophotometry (DTNB and released coenzyme A absorbance measurement, 412 nm) in the presence of organic solvent

Ethanol

30 % v/v

100

Digital

Continuous

100 mM KOAc/100 mM MES buffer, 0.25 mM 5.5-dithio-bis-(2-nitrobenzoic acid)

5.5

45°C

0.25 mM Hydroxymethylglutaroyl coenzyme A

Coenzyme A, Mevalonate

0.5 mM NADPH

—

Classical aqueous control (in %) | Racemic substrate

Activity - Classical

Activity measured by absorbance spectrophotometry (DTNB and released coenzyme A absorbance measurement, 412 nm) in the presence of organic solvent

Dimethylformamide (DMF)

20 % v/v

100

Digital

Continuous

100 mM KOAc/100 mM MES buffer, 0.25 mM 5.5-dithio-bis-(2-nitrobenzoic acid)

5.5

45°C

0.25 mM Hydroxymethylglutaroyl coenzyme A

Coenzyme A, Mevalonate

0.5 mM NADPH

—

Classical aqueous control (in %) | Racemic substrate

Activity - Classical

Activity measured by absorbance spectrophotometry (DTNB and released coenzyme A absorbance measurement, 412 nm) in the presence of organic solvent

Dimethylformamide (DMF)

15 % v/v

100

Digital

Continuous

100 mM KOAc/100 mM MES buffer, 0.25 mM 5.5-dithio-bis-(2-nitrobenzoic acid)

5.5

45°C

0.25 mM Hydroxymethylglutaroyl coenzyme A

Coenzyme A, Mevalonate

0.5 mM NADPH

—

Classical aqueous control (in %) | Racemic substrate

Activity - Classical

Activity measured by absorbance spectrophotometry (DTNB and released coenzyme A absorbance measurement, 412 nm) in the presence of organic solvent

Dimethylformamide (DMF)

10 % v/v

100

Digital

Continuous

100 mM KOAc/100 mM MES buffer, 0.25 mM 5.5-dithio-bis-(2-nitrobenzoic acid)

5.5

45°C

0.25 mM Hydroxymethylglutaroyl coenzyme A

Coenzyme A, Mevalonate

0.5 mM NADPH

—

Classical aqueous control (in %) | Racemic substrate

Activity - Classical

Activity measured by absorbance spectrophotometry (DTNB and released coenzyme A absorbance measurement, 412 nm) in the presence of organic solvent

Dimethylformamide (DMF)

5 % v/v

100

Digital

Continuous

100 mM KOAc/100 mM MES buffer, 0.25 mM 5.5-dithio-bis-(2-nitrobenzoic acid)

5.5

45°C

0.25 mM Hydroxymethylglutaroyl coenzyme A

Coenzyme A, Mevalonate

0.5 mM NADPH

—

Classical aqueous control (in %) | Racemic substrate

Activity - Classical

Activity measured by absorbance spectrophotometry (DTNB and released coenzyme A absorbance measurement, 412 nm) in the presence of organic solvent

Dimethylformamide (DMF)

2.5 % v/v

100

Digital

Continuous

100 mM KOAc/100 mM MES buffer, 0.25 mM 5.5-dithio-bis-(2-nitrobenzoic acid)

5.5

45°C

0.25 mM Hydroxymethylglutaroyl coenzyme A

Coenzyme A, Mevalonate

0.5 mM NADPH

—

Classical aqueous control (in %) | Racemic substrate

Activity - Classical

Activity measured by absorbance spectrophotometry (DTNB and released coenzyme A absorbance measurement, 412 nm) in the presence of organic solvent

Dimethylformamide (DMF)

1 % v/v

100

Digital

Continuous

100 mM KOAc/100 mM MES buffer, 0.25 mM 5.5-dithio-bis-(2-nitrobenzoic acid)

5.5

45°C

0.25 mM Hydroxymethylglutaroyl coenzyme A

Coenzyme A, Mevalonate

0.5 mM NADPH

—

Classical aqueous control (in %) | Racemic substrate

Activity - Classical

Activity measured by absorbance spectrophotometry (DTNB and released coenzyme A absorbance measurement, 412 nm) in the presence of organic solvent

Ethanol

60 % v/v

100

Digital

Continuous

100 mM KOAc/100 mM MES buffer, 0.25 mM 5.5-dithio-bis-(2-nitrobenzoic acid)

5.5

45°C

0.25 mM Hydroxymethylglutaroyl coenzyme A

Coenzyme A, Mevalonate

0.5 mM NADPH

—

Classical aqueous control (in %) | Racemic substrate

Activity - Classical

Activity measured by absorbance spectrophotometry (DTNB and released coenzyme A absorbance measurement, 412 nm) in the presence of organic solvent

Ethanol

50 % v/v

100

Digital

Continuous

100 mM KOAc/100 mM MES buffer, 0.25 mM 5.5-dithio-bis-(2-nitrobenzoic acid)

5.5

45°C

0.25 mM Hydroxymethylglutaroyl coenzyme A

Coenzyme A, Mevalonate

0.5 mM NADPH

—

Classical aqueous control (in %) | Racemic substrate

Activity - Classical

Activity measured by absorbance spectrophotometry (DTNB and released coenzyme A absorbance measurement, 412 nm) in the presence of organic solvent

Ethanol

40 % v/v

100

Digital

Continuous

100 mM KOAc/100 mM MES buffer, 0.25 mM 5.5-dithio-bis-(2-nitrobenzoic acid)

5.5

45°C

0.25 mM Hydroxymethylglutaroyl coenzyme A

Coenzyme A, Mevalonate

0.5 mM NADPH

—

Classical aqueous control (in %) | Racemic substrate

Activity - Classical

Activity measured by absorbance spectrophotometry (DTNB and released coenzyme A absorbance measurement, 412 nm) in the presence of organic solvent

Dimethylformamide (DMF)

30 % v/v

100

Digital

Continuous

100 mM KOAc/100 mM MES buffer, 0.25 mM 5.5-dithio-bis-(2-nitrobenzoic acid)

5.5

45°C

0.25 mM Hydroxymethylglutaroyl coenzyme A

Coenzyme A, Mevalonate

0.5 mM NADPH

—

Classical aqueous control (in %) | Racemic substrate

Activity - Classical

Activity measured by absorbance spectrophotometry (DTNB and released coenzyme A absorbance measurement, 412 nm) in the presence of organic solvent

Ethanol

20 % v/v

100

Digital

Continuous

100 mM KOAc/100 mM MES buffer, 0.25 mM 5.5-dithio-bis-(2-nitrobenzoic acid)

5.5

45°C

0.25 mM Hydroxymethylglutaroyl coenzyme A

Coenzyme A, Mevalonate

0.5 mM NADPH

—

Classical aqueous control (in %) | Racemic substrate

Activity - Classical

Activity measured by absorbance spectrophotometry (DTNB and released coenzyme A absorbance measurement, 412 nm) in the presence of organic solvent

Ethanol

15 % v/v

100

Digital

Continuous

100 mM KOAc/100 mM MES buffer, 0.25 mM 5.5-dithio-bis-(2-nitrobenzoic acid)

5.5

45°C

0.25 mM Hydroxymethylglutaroyl coenzyme A

Coenzyme A, Mevalonate

0.5 mM NADPH

—

Classical aqueous control (in %) | Racemic substrate

Activity - Classical

Activity measured by absorbance spectrophotometry (DTNB and released coenzyme A absorbance measurement, 412 nm) in the presence of organic solvent

Ethanol

10 % v/v

100

Digital

Continuous

100 mM KOAc/100 mM MES buffer, 0.25 mM 5.5-dithio-bis-(2-nitrobenzoic acid)

5.5

45°C

0.25 mM Hydroxymethylglutaroyl coenzyme A

Coenzyme A, Mevalonate

0.5 mM NADPH

—

Classical aqueous control (in %) | Racemic substrate

Activity - Classical

Activity measured by absorbance spectrophotometry (DTNB and released coenzyme A absorbance measurement, 412 nm) in the presence of organic solvent

Ethanol

5 % v/v

100

Digital

Continuous

100 mM KOAc/100 mM MES buffer, 0.25 mM 5.5-dithio-bis-(2-nitrobenzoic acid)

5.5

45°C

0.25 mM Hydroxymethylglutaroyl coenzyme A

Coenzyme A, Mevalonate

0.5 mM NADPH

—

Classical aqueous control (in %) | Racemic substrate

Activity - Classical

Activity measured by absorbance spectrophotometry (DTNB and released coenzyme A absorbance measurement, 412 nm) in the presence of organic solvent

Ethanol

2.5 % v/v

100

Digital

Continuous

100 mM KOAc/100 mM MES buffer, 0.25 mM 5.5-dithio-bis-(2-nitrobenzoic acid)

5.5

45°C

0.25 mM Hydroxymethylglutaroyl coenzyme A

Coenzyme A, Mevalonate

0.5 mM NADPH

—

Classical aqueous control (in %) | Racemic substrate

Activity - Classical

Activity measured by absorbance spectrophotometry (DTNB and released coenzyme A absorbance measurement, 412 nm) in the presence of organic solvent

Methanol

60 % v/v

100

Digital

Continuous

100 mM KOAc/100 mM MES buffer, 0.25 mM 5.5-dithio-bis-(2-nitrobenzoic acid)

5.5

45°C

0.25 mM Hydroxymethylglutaroyl coenzyme A

Coenzyme A, Mevalonate

0.5 mM NADPH

—

Classical aqueous control (in %) | Racemic substrate

Activity - Classical

Activity measured by absorbance spectrophotometry (DTNB and released coenzyme A absorbance measurement, 412 nm) in the presence of organic solvent

Methanol

50 % v/v

100

Digital

Continuous

100 mM KOAc/100 mM MES buffer, 0.25 mM 5.5-dithio-bis-(2-nitrobenzoic acid)

5.5

45°C

0.25 mM Hydroxymethylglutaroyl coenzyme A

Coenzyme A, Mevalonate

0.5 mM NADPH

—

Classical aqueous control (in %) | Racemic substrate

Activity - Classical

Activity measured by absorbance spectrophotometry (DTNB and released coenzyme A absorbance measurement, 412 nm) in the presence of organic solvent

Methanol

40 % v/v

100

Digital

Continuous

100 mM KOAc/100 mM MES buffer, 0.25 mM 5.5-dithio-bis-(2-nitrobenzoic acid)

5.5

45°C

0.25 mM Hydroxymethylglutaroyl coenzyme A

Coenzyme A, Mevalonate

0.5 mM NADPH

—

Classical aqueous control (in %) | Racemic substrate

Activity - Classical

Activity measured by absorbance spectrophotometry (DTNB and released coenzyme A absorbance measurement, 412 nm) in the presence of organic solvent

Methanol

30 % v/v

100

Digital

Continuous

100 mM KOAc/100 mM MES buffer, 0.25 mM 5.5-dithio-bis-(2-nitrobenzoic acid)

5.5

45°C

0.25 mM Hydroxymethylglutaroyl coenzyme A

Coenzyme A, Mevalonate

0.5 mM NADPH

—

Classical aqueous control (in %) | Racemic substrate

Activity - Classical

Activity measured by absorbance spectrophotometry (DTNB and released coenzyme A absorbance measurement, 412 nm) in the presence of organic solvent

Isopropanol

40 % v/v

100

Digital

Continuous

100 mM KOAc/100 mM MES buffer, 0.25 mM 5.5-dithio-bis-(2-nitrobenzoic acid)

5.5

45°C

0.25 mM Hydroxymethylglutaroyl coenzyme A

Coenzyme A, Mevalonate

0.5 mM NADPH

—

Classical aqueous control (in %) | Racemic substrate

Activity - Classical

Activity measured by absorbance spectrophotometry (DTNB and released coenzyme A absorbance measurement, 412 nm) in the presence of organic solvent

Acetone

60 % v/v

100

Digital

Continuous

100 mM KOAc/100 mM MES buffer, 0.25 mM 5.5-dithio-bis-(2-nitrobenzoic acid)

5.5

45°C

0.25 mM Hydroxymethylglutaroyl coenzyme A

Coenzyme A, Mevalonate

0.5 mM NADPH

—

Classical aqueous control (in %) | Racemic substrate

Activity - Classical

Activity measured by absorbance spectrophotometry (DTNB and released coenzyme A absorbance measurement, 412 nm) in the presence of organic solvent

Acetone

50 % v/v

100

Digital

Continuous

100 mM KOAc/100 mM MES buffer, 0.25 mM 5.5-dithio-bis-(2-nitrobenzoic acid)

5.5

45°C

0.25 mM Hydroxymethylglutaroyl coenzyme A

Coenzyme A, Mevalonate

0.5 mM NADPH

—

Classical aqueous control (in %) | Racemic substrate

Activity - Classical

Activity measured by absorbance spectrophotometry (DTNB and released coenzyme A absorbance measurement, 412 nm) in the presence of organic solvent

Acetone

40 % v/v

100

Digital

Continuous

100 mM KOAc/100 mM MES buffer, 0.25 mM 5.5-dithio-bis-(2-nitrobenzoic acid)

5.5

45°C

0.25 mM Hydroxymethylglutaroyl coenzyme A

Coenzyme A, Mevalonate

0.5 mM NADPH

—

Classical aqueous control (in %) | Racemic substrate

Activity - Classical

Activity measured by absorbance spectrophotometry (DTNB and released coenzyme A absorbance measurement, 412 nm) in the presence of organic solvent

Acetone

30 % v/v

100

Digital

Continuous

100 mM KOAc/100 mM MES buffer, 0.25 mM 5.5-dithio-bis-(2-nitrobenzoic acid)

5.5

45°C

0.25 mM Hydroxymethylglutaroyl coenzyme A

Coenzyme A, Mevalonate

0.5 mM NADPH

—

Classical aqueous control (in %) | Racemic substrate

Activity - Classical

Activity measured by absorbance spectrophotometry (DTNB and released coenzyme A absorbance measurement, 412 nm) in the presence of organic solvent

Acetone

20 % v/v

100

Digital

Continuous

100 mM KOAc/100 mM MES buffer, 0.25 mM 5.5-dithio-bis-(2-nitrobenzoic acid)

5.5

45°C

0.25 mM Hydroxymethylglutaroyl coenzyme A

Coenzyme A, Mevalonate

0.5 mM NADPH

—

Classical aqueous control (in %) | Racemic substrate

Activity - Classical

Activity measured by absorbance spectrophotometry (DTNB and released coenzyme A absorbance measurement, 412 nm) in the presence of organic solvent

Acetone

10 % v/v

100

Digital

Continuous

100 mM KOAc/100 mM MES buffer, 0.25 mM 5.5-dithio-bis-(2-nitrobenzoic acid)

5.5

45°C

0.25 mM Hydroxymethylglutaroyl coenzyme A

Coenzyme A, Mevalonate

0.5 mM NADPH

—

Classical aqueous control (in %) | Racemic substrate

Activity - Classical

Activity measured by absorbance spectrophotometry (DTNB and released coenzyme A absorbance measurement, 412 nm) in the presence of organic solvent

Acetone

5 % v/v

100

Digital

Continuous

100 mM KOAc/100 mM MES buffer, 0.25 mM 5.5-dithio-bis-(2-nitrobenzoic acid)

5.5

45°C

0.25 mM Hydroxymethylglutaroyl coenzyme A

Coenzyme A, Mevalonate

0.5 mM NADPH

—

Classical aqueous control (in %) | Racemic substrate

Activity - Classical

Activity measured by absorbance spectrophotometry (DTNB and released coenzyme A absorbance measurement, 412 nm) in the presence of organic solvent

Isopropanol

60 % v/v

100

Digital

Continuous

100 mM KOAc/100 mM MES buffer, 0.25 mM 5.5-dithio-bis-(2-nitrobenzoic acid)

5.5

45°C

0.25 mM Hydroxymethylglutaroyl coenzyme A

Coenzyme A, Mevalonate

0.5 mM NADPH

—

Classical aqueous control (in %) | Racemic substrate

Activity - Classical

Activity measured by absorbance spectrophotometry (DTNB and released coenzyme A absorbance measurement, 412 nm) in the presence of organic solvent

Isopropanol

50 % v/v

100

Digital

Continuous

100 mM KOAc/100 mM MES buffer, 0.25 mM 5.5-dithio-bis-(2-nitrobenzoic acid)

5.5

45°C

0.25 mM Hydroxymethylglutaroyl coenzyme A

Coenzyme A, Mevalonate

0.5 mM NADPH

—

Classical aqueous control (in %) | Racemic substrate

Activity - Classical

Activity measured by absorbance spectrophotometry (DTNB and released coenzyme A absorbance measurement, 412 nm) in the presence of organic solvent

Methanol

20 % v/v

100

Digital

Continuous

100 mM KOAc/100 mM MES buffer, 0.25 mM 5.5-dithio-bis-(2-nitrobenzoic acid)

5.5

45°C

0.25 mM Hydroxymethylglutaroyl coenzyme A

Coenzyme A, Mevalonate

0.5 mM NADPH

—

Classical aqueous control (in %) | Racemic substrate

Activity - Classical

Activity measured by absorbance spectrophotometry (DTNB and released coenzyme A absorbance measurement, 412 nm) in the presence of organic solvent

Isopropanol

30 % v/v

100

Digital

Continuous

100 mM KOAc/100 mM MES buffer, 0.25 mM 5.5-dithio-bis-(2-nitrobenzoic acid)

5.5

45°C

0.25 mM Hydroxymethylglutaroyl coenzyme A

Coenzyme A, Mevalonate

0.5 mM NADPH

—

Classical aqueous control (in %) | Racemic substrate

Activity - Classical

Activity measured by absorbance spectrophotometry (DTNB and released coenzyme A absorbance measurement, 412 nm) in the presence of organic solvent

Isopropanol

20 % v/v

100

Digital

Continuous

100 mM KOAc/100 mM MES buffer, 0.25 mM 5.5-dithio-bis-(2-nitrobenzoic acid)

5.5

45°C

0.25 mM Hydroxymethylglutaroyl coenzyme A

Coenzyme A, Mevalonate

0.5 mM NADPH

—

Classical aqueous control (in %) | Racemic substrate

Activity - Classical

Activity measured by absorbance spectrophotometry (DTNB and released coenzyme A absorbance measurement, 412 nm) in the presence of organic solvent

Isopropanol

15 % v/v

100

Digital

Continuous

100 mM KOAc/100 mM MES buffer, 0.25 mM 5.5-dithio-bis-(2-nitrobenzoic acid)

5.5

45°C

0.25 mM Hydroxymethylglutaroyl coenzyme A

Coenzyme A, Mevalonate

0.5 mM NADPH

—

Classical aqueous control (in %) | Racemic substrate

Activity - Classical

Activity measured by absorbance spectrophotometry (DTNB and released coenzyme A absorbance measurement, 412 nm) in the presence of organic solvent

Isopropanol

10 % v/v

100

Digital

Continuous

100 mM KOAc/100 mM MES buffer, 0.25 mM 5.5-dithio-bis-(2-nitrobenzoic acid)

5.5

45°C

0.25 mM Hydroxymethylglutaroyl coenzyme A

Coenzyme A, Mevalonate

0.5 mM NADPH

—

Classical aqueous control (in %) | Racemic substrate

Activity - Classical

Activity measured by absorbance spectrophotometry (DTNB and released coenzyme A absorbance measurement, 412 nm) in the presence of organic solvent

Isopropanol

5 % v/v

100

Digital

Continuous

100 mM KOAc/100 mM MES buffer, 0.25 mM 5.5-dithio-bis-(2-nitrobenzoic acid)

5.5

45°C

0.25 mM Hydroxymethylglutaroyl coenzyme A

Coenzyme A, Mevalonate

0.5 mM NADPH

—

Classical aqueous control (in %) | Racemic substrate

Activity - Classical

Activity measured by absorbance spectrophotometry (DTNB and released coenzyme A absorbance measurement, 412 nm) in the presence of organic solvent

Isopropanol

2.5 % v/v

100

Digital

Continuous

100 mM KOAc/100 mM MES buffer, 0.25 mM 5.5-dithio-bis-(2-nitrobenzoic acid)

5.5

45°C

0.25 mM Hydroxymethylglutaroyl coenzyme A

Coenzyme A, Mevalonate

0.5 mM NADPH

—

Classical aqueous control (in %) | Racemic substrate

Activity - Classical

Activity measured by absorbance spectrophotometry (DTNB and released coenzyme A absorbance measurement, 412 nm) in the presence of organic solvent

Dimethylformamide (DMF)

60 % v/v

100

Digital

Continuous

100 mM KOAc/100 mM MES buffer, 0.25 mM 5.5-dithio-bis-(2-nitrobenzoic acid)

5.5

45°C

0.25 mM Hydroxymethylglutaroyl coenzyme A

Coenzyme A, Mevalonate

0.5 mM NADPH

—

Classical aqueous control (in %) | Racemic substrate

Activity - Classical

Activity measured by absorbance spectrophotometry (DTNB and released coenzyme A absorbance measurement, 412 nm) in the presence of organic solvent

Dimethylformamide (DMF)

50 % v/v

100

Digital

Continuous

100 mM KOAc/100 mM MES buffer, 0.25 mM 5.5-dithio-bis-(2-nitrobenzoic acid)

5.5

45°C

0.25 mM Hydroxymethylglutaroyl coenzyme A

Coenzyme A, Mevalonate

0.5 mM NADPH

—

Classical aqueous control (in %) | Racemic substrate

Activity - Classical

Activity measured by absorbance spectrophotometry (DTNB and released coenzyme A absorbance measurement, 412 nm) in the presence of organic solvent

Dimethylformamide (DMF)

40 % v/v

100

Digital

Continuous

100 mM KOAc/100 mM MES buffer, 0.25 mM 5.5-dithio-bis-(2-nitrobenzoic acid)

5.5

45°C

0.25 mM Hydroxymethylglutaroyl coenzyme A

Coenzyme A, Mevalonate

0.5 mM NADPH

—

Classical aqueous control (in %) | Racemic substrate

Activity - Classical

Activity measured by absorbance spectrophotometry (DTNB and released coenzyme A absorbance measurement, 412 nm) in the presence of organic solvent

Tetrahydrofuran (THF)

5 % v/v

100

109

Digital

Continuous

100 mM KOAc/100 mM MES buffer, 0.25 mM 5.5-dithio-bis-(2-nitrobenzoic acid)

5.5

45°C

0.25 mM Hydroxymethylglutaroyl coenzyme A

Coenzyme A, Mevalonate

0.5 mM NADPH

—

Classical aqueous control (in %) | Racemic substrate

Activity - Classical

Activity measured by absorbance spectrophotometry (DTNB and released coenzyme A absorbance measurement, 412 nm) in the presence of organic solvent

Dimethyl Sulfoxide (DMSO)

5 % v/v

100

Digital

Continuous

100 mM KOAc/100 mM MES buffer, 0.25 mM 5.5-dithio-bis-(2-nitrobenzoic acid)

5.5

45°C

0.25 mM Hydroxymethylglutaroyl coenzyme A

Coenzyme A, Mevalonate

0.5 mM NADPH

—

Classical aqueous control (in %) | Racemic substrate

Activity - Classical

Activity measured by absorbance spectrophotometry (DTNB and released coenzyme A absorbance measurement, 412 nm) in the presence of organic solvent

Dimethyl Sulfoxide (DMSO)

2.5 % v/v

100

105

Digital

Continuous

100 mM KOAc/100 mM MES buffer, 0.25 mM 5.5-dithio-bis-(2-nitrobenzoic acid)

5.5

45°C

0.25 mM Hydroxymethylglutaroyl coenzyme A

Coenzyme A, Mevalonate

0.5 mM NADPH

—

Classical aqueous control (in %) | Racemic substrate

Activity - Classical

Activity measured by absorbance spectrophotometry (DTNB and released coenzyme A absorbance measurement, 412 nm) in the presence of organic solvent

Dimethyl Sulfoxide (DMSO)

1 % v/v

100

110

Digital

Continuous

100 mM KOAc/100 mM MES buffer, 0.25 mM 5.5-dithio-bis-(2-nitrobenzoic acid)

5.5

45°C

0.25 mM Hydroxymethylglutaroyl coenzyme A

Coenzyme A, Mevalonate

0.5 mM NADPH

—

Classical aqueous control (in %) | Racemic substrate

Activity - Classical

Activity measured by absorbance spectrophotometry (DTNB and released coenzyme A absorbance measurement, 412 nm) in the presence of organic solvent

Tetrahydrofuran (THF)

60 % v/v

100

Digital

Continuous

100 mM KOAc/100 mM MES buffer, 0.25 mM 5.5-dithio-bis-(2-nitrobenzoic acid)

5.5

45°C

0.25 mM Hydroxymethylglutaroyl coenzyme A

Coenzyme A, Mevalonate

0.5 mM NADPH

—

Classical aqueous control (in %) | Racemic substrate

Activity - Classical

Activity measured by absorbance spectrophotometry (DTNB and released coenzyme A absorbance measurement, 412 nm) in the presence of organic solvent

Tetrahydrofuran (THF)

50 % v/v

100

Digital

Continuous

100 mM KOAc/100 mM MES buffer, 0.25 mM 5.5-dithio-bis-(2-nitrobenzoic acid)

5.5

45°C

0.25 mM Hydroxymethylglutaroyl coenzyme A

Coenzyme A, Mevalonate

0.5 mM NADPH

—

Classical aqueous control (in %) | Racemic substrate

Activity - Classical

Activity measured by absorbance spectrophotometry (DTNB and released coenzyme A absorbance measurement, 412 nm) in the presence of organic solvent

Tetrahydrofuran (THF)

40 % v/v

100

Digital

Continuous

100 mM KOAc/100 mM MES buffer, 0.25 mM 5.5-dithio-bis-(2-nitrobenzoic acid)

5.5

45°C

0.25 mM Hydroxymethylglutaroyl coenzyme A

Coenzyme A, Mevalonate

0.5 mM NADPH

—

Classical aqueous control (in %) | Racemic substrate

Activity - Classical

Activity measured by absorbance spectrophotometry (DTNB and released coenzyme A absorbance measurement, 412 nm) in the presence of organic solvent

Tetrahydrofuran (THF)

30 % v/v

100

Digital

Continuous

100 mM KOAc/100 mM MES buffer, 0.25 mM 5.5-dithio-bis-(2-nitrobenzoic acid)

5.5

45°C

0.25 mM Hydroxymethylglutaroyl coenzyme A

Coenzyme A, Mevalonate

0.5 mM NADPH

—

Classical aqueous control (in %) | Racemic substrate

Activity - Classical

Activity measured by absorbance spectrophotometry (DTNB and released coenzyme A absorbance measurement, 412 nm) in the presence of organic solvent

Tetrahydrofuran (THF)

20 % v/v

100

Digital

Continuous

100 mM KOAc/100 mM MES buffer, 0.25 mM 5.5-dithio-bis-(2-nitrobenzoic acid)

5.5

45°C

0.25 mM Hydroxymethylglutaroyl coenzyme A

Coenzyme A, Mevalonate

0.5 mM NADPH

—

Classical aqueous control (in %) | Racemic substrate

Activity - Classical

Activity measured by absorbance spectrophotometry (DTNB and released coenzyme A absorbance measurement, 412 nm) in the presence of organic solvent

Tetrahydrofuran (THF)

10 % v/v

100

107

Digital

Continuous

100 mM KOAc/100 mM MES buffer, 0.25 mM 5.5-dithio-bis-(2-nitrobenzoic acid)

5.5

45°C

0.25 mM Hydroxymethylglutaroyl coenzyme A

Coenzyme A, Mevalonate

0.5 mM NADPH

—

Classical aqueous control (in %) | Racemic substrate

Activity - Classical

Activity measured by absorbance spectrophotometry (DTNB and released coenzyme A absorbance measurement, 412 nm) in the presence of organic solvent

Dimethyl Sulfoxide (DMSO)

10 % v/v

100

Digital

Continuous

100 mM KOAc/100 mM MES buffer, 0.25 mM 5.5-dithio-bis-(2-nitrobenzoic acid)

5.5

45°C

0.25 mM Hydroxymethylglutaroyl coenzyme A

Coenzyme A, Mevalonate

0.5 mM NADPH

—

Classical aqueous control (in %) | Racemic substrate

Activity - Classical

Activity measured by absorbance spectrophotometry (DTNB and released coenzyme A absorbance measurement, 412 nm) in the presence of organic solvent

Tetrahydrofuran (THF)

2.5 % v/v

100

106

Digital

Continuous

100 mM KOAc/100 mM MES buffer, 0.25 mM 5.5-dithio-bis-(2-nitrobenzoic acid)

5.5

45°C

0.25 mM Hydroxymethylglutaroyl coenzyme A

Coenzyme A, Mevalonate

0.5 mM NADPH

—

Classical aqueous control (in %) | Racemic substrate

Activity - Classical

Activity measured by absorbance spectrophotometry (DTNB and released coenzyme A absorbance measurement, 412 nm) in the presence of organic solvent

Acetonitrile

60 % v/v

100

Digital

Continuous

100 mM KOAc/100 mM MES buffer, 0.25 mM 5.5-dithio-bis-(2-nitrobenzoic acid)

5.5

45°C

0.25 mM Hydroxymethylglutaroyl coenzyme A

Coenzyme A, Mevalonate

0.5 mM NADPH

—

Classical aqueous control (in %) | Racemic substrate

Activity - Classical

Activity measured by absorbance spectrophotometry (DTNB and released coenzyme A absorbance measurement, 412 nm) in the presence of organic solvent

Acetonitrile

50 % v/v

100

Digital

Continuous

100 mM KOAc/100 mM MES buffer, 0.25 mM 5.5-dithio-bis-(2-nitrobenzoic acid)

5.5

45°C

0.25 mM Hydroxymethylglutaroyl coenzyme A

Coenzyme A, Mevalonate

0.5 mM NADPH

—

Classical aqueous control (in %) | Racemic substrate

Activity - Classical

Activity measured by absorbance spectrophotometry (DTNB and released coenzyme A absorbance measurement, 412 nm) in the presence of organic solvent

Acetonitrile

40 % v/v

100

Digital

Continuous

100 mM KOAc/100 mM MES buffer, 0.25 mM 5.5-dithio-bis-(2-nitrobenzoic acid)

5.5

45°C

0.25 mM Hydroxymethylglutaroyl coenzyme A

Coenzyme A, Mevalonate

0.5 mM NADPH

—

Classical aqueous control (in %) | Racemic substrate

Activity - Classical

Activity measured by absorbance spectrophotometry (DTNB and released coenzyme A absorbance measurement, 412 nm) in the presence of organic solvent

Acetonitrile

30 % v/v

100

Digital

Continuous

100 mM KOAc/100 mM MES buffer, 0.25 mM 5.5-dithio-bis-(2-nitrobenzoic acid)

5.5

45°C

0.25 mM Hydroxymethylglutaroyl coenzyme A

Coenzyme A, Mevalonate

0.5 mM NADPH

—

Classical aqueous control (in %) | Racemic substrate

Activity - Classical

Activity measured by absorbance spectrophotometry (DTNB and released coenzyme A absorbance measurement, 412 nm) in the presence of organic solvent

Acetonitrile

20 % v/v

100

101

Digital

Continuous

100 mM KOAc/100 mM MES buffer, 0.25 mM 5.5-dithio-bis-(2-nitrobenzoic acid)

5.5

45°C

0.25 mM Hydroxymethylglutaroyl coenzyme A

Coenzyme A, Mevalonate

0.5 mM NADPH

—

Classical aqueous control (in %) | Racemic substrate

Activity - Classical

Activity measured by absorbance spectrophotometry (DTNB and released coenzyme A absorbance measurement, 412 nm) in the presence of organic solvent

Acetonitrile

15 % v/v

100

106

Digital

Continuous

100 mM KOAc/100 mM MES buffer, 0.25 mM 5.5-dithio-bis-(2-nitrobenzoic acid)

5.5

45°C

0.25 mM Hydroxymethylglutaroyl coenzyme A

Coenzyme A, Mevalonate

0.5 mM NADPH

—

Classical aqueous control (in %) | Racemic substrate

Activity - Classical

Activity measured by absorbance spectrophotometry (DTNB and released coenzyme A absorbance measurement, 412 nm) in the presence of organic solvent

Acetonitrile

10 % v/v

100

117

Digital

Continuous

100 mM KOAc/100 mM MES buffer, 0.25 mM 5.5-dithio-bis-(2-nitrobenzoic acid)

5.5

45°C

0.25 mM Hydroxymethylglutaroyl coenzyme A

Coenzyme A, Mevalonate

0.5 mM NADPH

—

Classical aqueous control (in %) | Racemic substrate

Activity - Classical

Activity measured by absorbance spectrophotometry (DTNB and released coenzyme A absorbance measurement, 412 nm) in the presence of organic solvent

Acetonitrile

5 % v/v

100

111

Digital

Continuous

100 mM KOAc/100 mM MES buffer, 0.25 mM 5.5-dithio-bis-(2-nitrobenzoic acid)

5.5

45°C

0.25 mM Hydroxymethylglutaroyl coenzyme A

Coenzyme A, Mevalonate

0.5 mM NADPH

—

Classical aqueous control (in %) | Racemic substrate

Activity - Classical

Activity measured by absorbance spectrophotometry (DTNB and released coenzyme A absorbance measurement, 412 nm) in the presence of organic solvent

1,4-Dioxane

10 % v/v

100

Digital

Continuous

100 mM KOAc/100 mM MES buffer, 0.25 mM 5.5-dithio-bis-(2-nitrobenzoic acid)

5.5

45°C

0.25 mM Hydroxymethylglutaroyl coenzyme A

Coenzyme A, Mevalonate

0.5 mM NADPH

—

Classical aqueous control (in %) | Racemic substrate

Activity - Classical

Activity measured by absorbance spectrophotometry (DTNB and released coenzyme A absorbance measurement, 412 nm) in the presence of organic solvent

Methanol

15 % v/v

100

Digital

Continuous

100 mM KOAc/100 mM MES buffer, 0.25 mM 5.5-dithio-bis-(2-nitrobenzoic acid)

5.5

45°C

0.25 mM Hydroxymethylglutaroyl coenzyme A

Coenzyme A, Mevalonate

0.5 mM NADPH

—

Classical aqueous control (in %) | Racemic substrate

Activity - Classical

Activity measured by absorbance spectrophotometry (DTNB and released coenzyme A absorbance measurement, 412 nm) in the presence of organic solvent

Methanol

10 % v/v

100

Digital

Continuous

100 mM KOAc/100 mM MES buffer, 0.25 mM 5.5-dithio-bis-(2-nitrobenzoic acid)

5.5

45°C

0.25 mM Hydroxymethylglutaroyl coenzyme A

Coenzyme A, Mevalonate

0.5 mM NADPH

—

Classical aqueous control (in %) | Racemic substrate

Activity - Classical

Activity measured by absorbance spectrophotometry (DTNB and released coenzyme A absorbance measurement, 412 nm) in the presence of organic solvent

Methanol

5 % v/v

100

Digital

Continuous

100 mM KOAc/100 mM MES buffer, 0.25 mM 5.5-dithio-bis-(2-nitrobenzoic acid)

5.5

45°C

0.25 mM Hydroxymethylglutaroyl coenzyme A

Coenzyme A, Mevalonate

0.5 mM NADPH

—

Classical aqueous control (in %) | Racemic substrate

Activity - Classical

Activity measured by absorbance spectrophotometry (DTNB and released coenzyme A absorbance measurement, 412 nm) in the presence of organic solvent

Methanol

2.5 % v/v

100

Digital

Continuous

100 mM KOAc/100 mM MES buffer, 0.25 mM 5.5-dithio-bis-(2-nitrobenzoic acid)

5.5

45°C

0.25 mM Hydroxymethylglutaroyl coenzyme A

Coenzyme A, Mevalonate

0.5 mM NADPH

—

Classical aqueous control (in %) | Racemic substrate

Activity - Classical

Activity measured by absorbance spectrophotometry (DTNB and released coenzyme A absorbance measurement, 412 nm) in the presence of organic solvent

1,4-Dioxane

60 % v/v

100

Digital

Continuous

100 mM KOAc/100 mM MES buffer, 0.25 mM 5.5-dithio-bis-(2-nitrobenzoic acid)

5.5

45°C

0.25 mM Hydroxymethylglutaroyl coenzyme A

Coenzyme A, Mevalonate

0.5 mM NADPH

—

Classical aqueous control (in %) | Racemic substrate

Activity - Classical

Activity measured by absorbance spectrophotometry (DTNB and released coenzyme A absorbance measurement, 412 nm) in the presence of organic solvent

1,4-Dioxane

50 % v/v

100

Digital

Continuous

100 mM KOAc/100 mM MES buffer, 0.25 mM 5.5-dithio-bis-(2-nitrobenzoic acid)

5.5

45°C

0.25 mM Hydroxymethylglutaroyl coenzyme A

Coenzyme A, Mevalonate

0.5 mM NADPH

—

Classical aqueous control (in %) | Racemic substrate

Activity - Classical

Activity measured by absorbance spectrophotometry (DTNB and released coenzyme A absorbance measurement, 412 nm) in the presence of organic solvent

1,4-Dioxane

40 % v/v

100

Digital

Continuous

100 mM KOAc/100 mM MES buffer, 0.25 mM 5.5-dithio-bis-(2-nitrobenzoic acid)

5.5

45°C

0.25 mM Hydroxymethylglutaroyl coenzyme A

Coenzyme A, Mevalonate

0.5 mM NADPH

—

Classical aqueous control (in %) | Racemic substrate

Activity - Classical

Activity measured by absorbance spectrophotometry (DTNB and released coenzyme A absorbance measurement, 412 nm) in the presence of organic solvent

1,4-Dioxane

30 % v/v

100

Digital

Continuous

100 mM KOAc/100 mM MES buffer, 0.25 mM 5.5-dithio-bis-(2-nitrobenzoic acid)

5.5

45°C

0.25 mM Hydroxymethylglutaroyl coenzyme A

Coenzyme A, Mevalonate

0.5 mM NADPH

—

Classical aqueous control (in %) | Racemic substrate

Activity - Classical

Activity measured by absorbance spectrophotometry (DTNB and released coenzyme A absorbance measurement, 412 nm) in the presence of organic solvent

1,4-Dioxane

20 % v/v

100

Digital

Continuous

100 mM KOAc/100 mM MES buffer, 0.25 mM 5.5-dithio-bis-(2-nitrobenzoic acid)

5.5

45°C

0.25 mM Hydroxymethylglutaroyl coenzyme A

Coenzyme A, Mevalonate

0.5 mM NADPH

—

Classical aqueous control (in %) | Racemic substrate

Activity - Classical

Activity measured by absorbance spectrophotometry (DTNB and released coenzyme A absorbance measurement, 412 nm) in the presence of organic solvent

Acetonitrile

2.5 % v/v

100

105

Digital

Continuous

100 mM KOAc/100 mM MES buffer, 0.25 mM 5.5-dithio-bis-(2-nitrobenzoic acid)

5.5

45°C

0.25 mM Hydroxymethylglutaroyl coenzyme A

Coenzyme A, Mevalonate

0.5 mM NADPH

—

Classical aqueous control (in %) | Racemic substrate

Activity - Classical

Activity measured by absorbance spectrophotometry (DTNB and released coenzyme A absorbance measurement, 412 nm) in the presence of organic solvent

1,4-Dioxane

5 % v/v

100

Digital

Continuous

100 mM KOAc/100 mM MES buffer, 0.25 mM 5.5-dithio-bis-(2-nitrobenzoic acid)

5.5

45°C

0.25 mM Hydroxymethylglutaroyl coenzyme A

Coenzyme A, Mevalonate

0.5 mM NADPH

—

Classical aqueous control (in %) | Racemic substrate

Activity - Classical

Activity measured by absorbance spectrophotometry (DTNB and released coenzyme A absorbance measurement, 412 nm) in the presence of organic solvent

1,4-Dioxane

2.5 % v/v

100

Digital

Continuous

100 mM KOAc/100 mM MES buffer, 0.25 mM 5.5-dithio-bis-(2-nitrobenzoic acid)

5.5

45°C

0.25 mM Hydroxymethylglutaroyl coenzyme A

Coenzyme A, Mevalonate

0.5 mM NADPH

—

Classical aqueous control (in %) | Racemic substrate

Activity - Classical

Activity measured by absorbance spectrophotometry (DTNB and released coenzyme A absorbance measurement, 412 nm) in the presence of organic solvent

Dimethyl Sulfoxide (DMSO)

60 % v/v

100

Digital

Continuous

100 mM KOAc/100 mM MES buffer, 0.25 mM 5.5-dithio-bis-(2-nitrobenzoic acid)

5.5

45°C

0.25 mM Hydroxymethylglutaroyl coenzyme A

Coenzyme A, Mevalonate

0.5 mM NADPH

—

Classical aqueous control (in %) | Racemic substrate

Activity - Classical

Activity measured by absorbance spectrophotometry (DTNB and released coenzyme A absorbance measurement, 412 nm) in the presence of organic solvent

Dimethyl Sulfoxide (DMSO)

50 % v/v

100

Digital

Continuous

100 mM KOAc/100 mM MES buffer, 0.25 mM 5.5-dithio-bis-(2-nitrobenzoic acid)

5.5

45°C

0.25 mM Hydroxymethylglutaroyl coenzyme A

Coenzyme A, Mevalonate

0.5 mM NADPH

—

Classical aqueous control (in %) | Racemic substrate

Activity - Classical

Activity measured by absorbance spectrophotometry (DTNB and released coenzyme A absorbance measurement, 412 nm) in the presence of organic solvent

Dimethyl Sulfoxide (DMSO)

40 % v/v

100

Digital

Continuous

100 mM KOAc/100 mM MES buffer, 0.25 mM 5.5-dithio-bis-(2-nitrobenzoic acid)

5.5

45°C

0.25 mM Hydroxymethylglutaroyl coenzyme A

Coenzyme A, Mevalonate

0.5 mM NADPH

—

Classical aqueous control (in %) | Racemic substrate

Activity - Classical

Activity measured by absorbance spectrophotometry (DTNB and released coenzyme A absorbance measurement, 412 nm) in the presence of organic solvent

Dimethyl Sulfoxide (DMSO)

30 % v/v

100

Digital

Continuous

100 mM KOAc/100 mM MES buffer, 0.25 mM 5.5-dithio-bis-(2-nitrobenzoic acid)

5.5

45°C

0.25 mM Hydroxymethylglutaroyl coenzyme A

Coenzyme A, Mevalonate

0.5 mM NADPH

—

Classical aqueous control (in %) | Racemic substrate

Activity - Classical

Activity measured by absorbance spectrophotometry (DTNB and released coenzyme A absorbance measurement, 412 nm) in the presence of organic solvent

Dimethyl Sulfoxide (DMSO)

20 % v/v

100

Digital

Continuous

100 mM KOAc/100 mM MES buffer, 0.25 mM 5.5-dithio-bis-(2-nitrobenzoic acid)

5.5

45°C

0.25 mM Hydroxymethylglutaroyl coenzyme A

Coenzyme A, Mevalonate

0.5 mM NADPH

—

Classical aqueous control (in %) | Racemic substrate

Activity - Classical

Activity measured by absorbance spectrophotometry (DTNB and released coenzyme A absorbance measurement, 412 nm) in the presence of organic solvent

Dimethyl Sulfoxide (DMSO)

15 % v/v

100

Digital

Continuous

100 mM KOAc/100 mM MES buffer, 0.25 mM 5.5-dithio-bis-(2-nitrobenzoic acid)

5.5

45°C

0.25 mM Hydroxymethylglutaroyl coenzyme A

Coenzyme A, Mevalonate

0.5 mM NADPH

—

Classical aqueous control (in %) | Racemic substrate

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Visualization : Activity — Classical

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

Structure

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3-hydroxy-3-methylglutaryl coenzyme A reductase from Sulfolobus solfataricus

Enzyme Description

Sequence

Experimental Data (79 measurements)

Visualization : Activity — Classical

Structure

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