Short-chain alcohol dehydrogenase from Thermococcus sibiricus

Enzyme Description

Species

Thermococcus sibiricus ↗

Extremophile

Yes "hyperthermophilic" organism

EC Number

1.1.1.1 ↗

Sequence

Length: 234 amino acids

MKVAVITGASRGIGEAIARALARDGYALALGARSVDRLEKIAHELMQEQGVEVFYHHLDVSKAESVEEFSKKVLERFGDVDVVVANAGLGYFKRLEELSEEEFHEMIEVNLLGVWRTLKAFLDSLKRTGGLALVTTSDVSARLIPYGGGYVSTKWAARALVRTFQIENPDVRFFELRPGAVDTYFGGSKPGKPKEKGYLKPDEIAEAVRCLLKLPKDVRVEELMLRSVYQRPEY

Tatiana N. Stekhanova et al. (2010) — Characterization of a Thermostable Short-Chain Alcohol Dehydrogenase from the Hyperthermophilic Archaeon Thermococcus sibiricus

Applied and Environmental Microbiology · doi:10.1128/AEM.02797-09 ↗ · Activity - Classical Stability - Incubation

24 measurements ▼

Database ID

UniProt: C6A190 ↗

Sequence Annotation

Explicit - Provided GenBank Accession Number

Protein Source

Recombinant, host bacterium Escherichia coli Rosetta-gami (DE3)

Experimental Data (24 measurements)

24 measurements — page 2 of 2

Property	Assay	Solvent	Solvent Volume	Aqueous Reference	Measured Value	Units	Solution	pH	Temperature	Substrate(s)	Product(s)	Cofactor(s)	Shaking	Comments
Stability - Incubation	Activity after incubation (4 hours at 55°C), measured by absorance spectrophotometry (change in NADPH absorbance measurement, 340 nm) in aqueous phase	Ethyl Acetate	50%	100	47	%	Incubation: 50 mM Gly-NaOH, Assay: 50 mM Gly-NaOH	Incubation: 10.5, Assay: 10.5	Incubation: 55°C, Assay: 60°C	250 mM Isopropanol	Propanone	NADP+	—	Water-incubated control (in %), assay in aqueous phase \| Uncertainty about whether activity was really measured in aqueous phase
Stability - Incubation	Activity after incubation (4 hours at 55°C), measured by absorance spectrophotometry (change in NADPH absorbance measurement, 340 nm) in aqueous phase	Acetonitrile	50%	100	95	%	Incubation: 50 mM Gly-NaOH, Assay: 50 mM Gly-NaOH	Incubation: 10.5, Assay: 10.5	Incubation: 55°C, Assay: 60°C	250 mM Isopropanol	Propanone	NADP+	—	Water-incubated control (in %), assay in aqueous phase \| Uncertainty about whether activity was really measured in aqueous phase
Stability - Incubation	Activity after incubation (4 hours at 55°C), measured by absorance spectrophotometry (change in NADPH absorbance measurement, 340 nm) in aqueous phase	Methanol	50%	100	98	%	Incubation: 50 mM Gly-NaOH, Assay: 50 mM Gly-NaOH	Incubation: 10.5, Assay: 10.5	Incubation: 55°C, Assay: 60°C	250 mM Isopropanol	Propanone	NADP+	—	Water-incubated control (in %), assay in aqueous phase \| Uncertainty about whether activity was really measured in aqueous phase
Stability - Incubation	Activity after incubation (4 hours at 55°C), measured by absorance spectrophotometry (change in NADPH absorbance measurement, 340 nm) in aqueous phase	Dimethylformamide (DMF)	50%	100	101	%	Incubation: 50 mM Gly-NaOH, Assay: 50 mM Gly-NaOH	Incubation: 10.5, Assay: 10.5	Incubation: 55°C, Assay: 60°C	250 mM Isopropanol	Propanone	NADP+	—	Water-incubated control (in %), assay in aqueous phase \| Uncertainty about whether activity was really measured in aqueous phase

Activity - Classical

Activity measured by absorbance spectrophotometry (change in NADPH absorbance measurement, 340 nm) in the presence of organic solvent

Ethyl Acetate

Saturation

100

50 mM Gly-NaOH buffer without NaCl

10.5

60°C

250 mM Isopropanol

Propanone

NADP+

—

Classical aqueous control (in %)

Activity - Classical

Activity measured by absorbance spectrophotometry (change in NADPH absorbance measurement, 340 nm) in the presence of organic solvent

n-Decane

Saturation

100

107

50 mM Gly-NaOH buffer with 600 mM NaCl

10.5

60°C

250 mM Isopropanol

Propanone

NADP+

—

Classical aqueous control (in %)

Activity - Classical

Activity measured by absorbance spectrophotometry (change in NADPH absorbance measurement, 340 nm) in the presence of organic solvent

n-Hexane

Saturation

100

118

50 mM Gly-NaOH buffer with 600 mM NaCl

10.5

60°C

250 mM Isopropanol

Propanone

NADP+

—

Classical aqueous control (in %)

Activity - Classical

Activity measured by absorbance spectrophotometry (change in NADPH absorbance measurement, 340 nm) in the presence of organic solvent

Chloroform

Saturation

100

50 mM Gly-NaOH buffer with 600 mM NaCl

10.5

60°C

250 mM Isopropanol

Propanone

NADP+

—

Classical aqueous control (in %)

Activity - Classical

Activity measured by absorbance spectrophotometry (change in NADPH absorbance measurement, 340 nm) in the presence of organic solvent

Ethyl Acetate

Saturation

100

50 mM Gly-NaOH buffer with 600 mM NaCl

10.5

60°C

250 mM Isopropanol

Propanone

NADP+

—

Classical aqueous control (in %)

Activity - Classical

Activity measured by absorbance spectrophotometry (change in NADPH absorbance measurement, 340 nm) in the presence of organic solvent

Acetonitrile

50%

100

50 mM Gly-NaOH buffer with 600 mM NaCl

10.5

60°C

250 mM Isopropanol

Propanone

NADP+

—

Classical aqueous control (in %)

Activity - Classical

Activity measured by absorbance spectrophotometry (change in NADPH absorbance measurement, 340 nm) in the presence of organic solvent

Methanol

50%

100

50 mM Gly-NaOH buffer with 600 mM NaCl

10.5

60°C

250 mM Isopropanol

Propanone

NADP+

—

Classical aqueous control (in %)

Activity - Classical

Activity measured by absorbance spectrophotometry (change in NADPH absorbance measurement, 340 nm) in the presence of organic solvent

Dimethylformamide (DMF)

50%

100

50 mM Gly-NaOH buffer with 600 mM NaCl

10.5

60°C

250 mM Isopropanol

Propanone

NADP+

—

Classical aqueous control (in %)

Activity - Classical

Activity measured by absorbance spectrophotometry (change in NADPH absorbance measurement, 340 nm) in the presence of organic solvent

Dimethyl Sulfoxide (DMSO)

50%

100

50 mM Gly-NaOH buffer with 600 mM NaCl

10.5

60°C

250 mM Isopropanol

Propanone

NADP+

—

Classical aqueous control (in %)

Activity - Classical

Activity measured by absorbance spectrophotometry (change in NADPH absorbance measurement, 340 nm) in the presence of organic solvent

n-Decane

Saturation

100

50 mM Gly-NaOH buffer without NaCl

10.5

60°C

250 mM Isopropanol

Propanone

NADP+

—

Classical aqueous control (in %)

Activity - Classical

Activity measured by absorbance spectrophotometry (change in NADPH absorbance measurement, 340 nm) in the presence of organic solvent

n-Hexane

Saturation

100

50 mM Gly-NaOH buffer without NaCl

10.5

60°C

250 mM Isopropanol

Propanone

NADP+

—

Classical aqueous control (in %)

Activity - Classical

Activity measured by absorbance spectrophotometry (change in NADPH absorbance measurement, 340 nm) in the presence of organic solvent

Chloroform

Saturation

100

50 mM Gly-NaOH buffer without NaCl

10.5

60°C

250 mM Isopropanol

Propanone

NADP+

—

Classical aqueous control (in %)

Activity - Classical

Activity measured by absorbance spectrophotometry (change in NADPH absorbance measurement, 340 nm) in the presence of organic solvent

Acetonitrile

50%

100

50 mM Gly-NaOH buffer without NaCl

10.5

60°C

250 mM Isopropanol

Propanone

NADP+

—

Classical aqueous control (in %)

Activity - Classical

Activity measured by absorbance spectrophotometry (change in NADPH absorbance measurement, 340 nm) in the presence of organic solvent

Methanol

50%

100

50 mM Gly-NaOH buffer without NaCl

10.5

60°C

250 mM Isopropanol

Propanone

NADP+

—

Classical aqueous control (in %)

Activity - Classical

Activity measured by absorbance spectrophotometry (change in NADPH absorbance measurement, 340 nm) in the presence of organic solvent

Dimethylformamide (DMF)

50%

100

50 mM Gly-NaOH buffer without NaCl

10.5

60°C

250 mM Isopropanol

Propanone

NADP+

—

Classical aqueous control (in %)

Activity - Classical

Activity measured by absorbance spectrophotometry (change in NADPH absorbance measurement, 340 nm) in the presence of organic solvent

Dimethyl Sulfoxide (DMSO)

50%

100

50 mM Gly-NaOH buffer without NaCl

10.5

60°C

250 mM Isopropanol

Propanone

NADP+

—

Classical aqueous control (in %)

Stability - Incubation

Activity after incubation (4 hours at 55°C), measured by absorance spectrophotometry (change in NADPH absorbance measurement, 340 nm) in aqueous phase

Dimethyl Sulfoxide (DMSO)

50%

100

Incubation: 50 mM Gly-NaOH, Assay: 50 mM Gly-NaOH

Incubation: 10.5, Assay: 10.5

Incubation: 55°C, Assay: 60°C

250 mM Isopropanol

Propanone

NADP+

—

Water-incubated control (in %), assay in aqueous phase | Uncertainty about whether activity was really measured in aqueous phase

Stability - Incubation

Activity after incubation (4 hours at 55°C), measured by absorance spectrophotometry (change in NADPH absorbance measurement, 340 nm) in aqueous phase

n-Decane

50%

100

Incubation: 50 mM Gly-NaOH, Assay: 50 mM Gly-NaOH

Incubation: 10.5, Assay: 10.5

Incubation: 55°C, Assay: 60°C

250 mM Isopropanol

Propanone

NADP+

—

Water-incubated control (in %), assay in aqueous phase | Uncertainty about whether activity was really measured in aqueous phase

Stability - Incubation

Activity after incubation (4 hours at 55°C), measured by absorance spectrophotometry (change in NADPH absorbance measurement, 340 nm) in aqueous phase

n-Hexane

50%

100

105

Incubation: 50 mM Gly-NaOH, Assay: 50 mM Gly-NaOH

Incubation: 10.5, Assay: 10.5

Incubation: 55°C, Assay: 60°C

250 mM Isopropanol

Propanone

NADP+

—

Water-incubated control (in %), assay in aqueous phase | Uncertainty about whether activity was really measured in aqueous phase

Stability - Incubation

Activity after incubation (4 hours at 55°C), measured by absorance spectrophotometry (change in NADPH absorbance measurement, 340 nm) in aqueous phase

Chloroform

50%

100

105

Incubation: 50 mM Gly-NaOH, Assay: 50 mM Gly-NaOH

Incubation: 10.5, Assay: 10.5

Incubation: 55°C, Assay: 60°C

250 mM Isopropanol

Propanone

NADP+

—

Water-incubated control (in %), assay in aqueous phase | Uncertainty about whether activity was really measured in aqueous phase

Stability - Incubation

Activity after incubation (4 hours at 55°C), measured by absorance spectrophotometry (change in NADPH absorbance measurement, 340 nm) in aqueous phase

Ethyl Acetate

50%

100

Incubation: 50 mM Gly-NaOH, Assay: 50 mM Gly-NaOH

Incubation: 10.5, Assay: 10.5

Incubation: 55°C, Assay: 60°C

250 mM Isopropanol

Propanone

NADP+

—

Water-incubated control (in %), assay in aqueous phase | Uncertainty about whether activity was really measured in aqueous phase

Stability - Incubation

Activity after incubation (4 hours at 55°C), measured by absorance spectrophotometry (change in NADPH absorbance measurement, 340 nm) in aqueous phase

Acetonitrile

50%

100

Incubation: 50 mM Gly-NaOH, Assay: 50 mM Gly-NaOH

Incubation: 10.5, Assay: 10.5

Incubation: 55°C, Assay: 60°C

250 mM Isopropanol

Propanone

NADP+

—

Water-incubated control (in %), assay in aqueous phase | Uncertainty about whether activity was really measured in aqueous phase

Stability - Incubation

Activity after incubation (4 hours at 55°C), measured by absorance spectrophotometry (change in NADPH absorbance measurement, 340 nm) in aqueous phase

Methanol

50%

100

Incubation: 50 mM Gly-NaOH, Assay: 50 mM Gly-NaOH

Incubation: 10.5, Assay: 10.5

Incubation: 55°C, Assay: 60°C

250 mM Isopropanol

Propanone

NADP+

—

Water-incubated control (in %), assay in aqueous phase | Uncertainty about whether activity was really measured in aqueous phase

Stability - Incubation

Activity after incubation (4 hours at 55°C), measured by absorance spectrophotometry (change in NADPH absorbance measurement, 340 nm) in aqueous phase

Dimethylformamide (DMF)

50%

100

101

Incubation: 50 mM Gly-NaOH, Assay: 50 mM Gly-NaOH

Incubation: 10.5, Assay: 10.5

Incubation: 55°C, Assay: 60°C

250 mM Isopropanol

Propanone

NADP+

—

Water-incubated control (in %), assay in aqueous phase | Uncertainty about whether activity was really measured in aqueous phase

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Visualization : Activity — Classical

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

Visualization : Stability — Incubation

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

Structure

🧬

No structure available for this protein.

Short-chain alcohol dehydrogenase from Thermococcus sibiricus

Enzyme Description

Sequence

Experimental Data (24 measurements)

Visualization : Activity — Classical

Visualization : Stability — Incubation

Structure

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