Transketolase (gene TKtmar) from Thermotoga maritima DSM3109

Enzyme Description

Species

Thermotoga maritima DSM3109 ↗

Extremophile

Yes "hyperthermophilic" organism

EC Number

2.2.1.1 ↗

Sequence

Length: 635 amino acids

MERFPYEKLPESELKELKELGRLCRGDILKMTYIANSGHPGGSMSSIDLYLTVFKYAKLRPVDDPARDRIVISHGHTSPGVYAAMARLGFVDLDEVLAGFRHPASVFEGHVTRGVGIIDWTTGNLGQGLSAGLGFALASRFTGKDYHVFVLMSDAEQAKGQVAEARRVAKKYGVTNLTVIIDYNDAQISGRARDVMPVNIKENYLADGWRVIEIDGHDYEQIYLALKEAVEDELNPVAIIAKTVMGKGVSFMENEVKYHGKPLNREELEKALAELGIENDVDVYIEKRKQLPVEKHKKVYKTYPIKIDTGEPITYTSPTDNRSAFGKAILDLVKKNVNNPETTPIVAVDCDLKGSVKLDLLDKEFPERLLEVGVQEHNAAAMAGALSAEGVITFFADFGVFGISETYNQHRLNAINGTNLKVVVTHCGLNVGEDGKTHHGLDYVSGPMNWYGFKVIVPGDPNQTDRVVRYAAKEYGNFVIAMGRSKLPIILDENGKPFFGEGYTFEYGKIDVVRKGDDAVIITYGSTLCEAVNAADELKKEGVNVAVLNVSCPVDLDIETLKMVDGKPVLVVEDHNVFTGLGSFLGTTLLENGIIPKKYVRVGVPEFAVSGSYTMLYKLYGLDKDGIISRLREML

Max Cárdenas-Fernández et al. (2021) — Characterisation of a hyperthermophilic transketolase from Thermotoga maritima DSM3109 as a biocatalyst for 7-keto-octuronic acid synthesis

Organic & Biomolecular Chemistry · doi:10.1039/d1ob01237a ↗ · Stability - Incubation

24 measurements ▼

Database ID

UniParc: UPI00000D3857 ↗

Sequence Annotation

Explicit - Provided GenBank Accession Number

Protein Source

Recombinant, host bacterium Escherichia coli BL21 (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 (2 hours at 50°C), measured by absorbance spectrophotometry (NADH absorbance measurement, as reaction product glyceraldehyde reacts with alcohol dehydrogenase, 340 nm) in aqueous phase	Methanol	10%	101.4	110.3	%	Incubation: 100 mM Tris-HCl buffer, Assay: 100 mM Tris-HCl buffer, 0.2 MM NADH, 25 U/ml alcohol dehydrogenase from Saccharomyces cerevisiae	Incubation: 7, Assay: 7	Incubation: 50°C, Assay: 40°C	100 mM D-Erythrose, 9.1 mM D-Ribose 5-phosphate	Glyceraldehyde , Sedoheptulose-7-phosphate	0.5 mM MgCl₂, 0.1 mM ThDP	—	Water-incubated control (in %), assay in aqueous phase \| Uncertainty about whether assay really was in aqueous phase
Stability - Incubation	Activity after incubation (30 hours at 50°C), measured by absorbance spectrophotometry (NADH absorbance measurement, as reaction product glyceraldehyde reacts with alcohol dehydrogenase, 340 nm) in aqueous phase	Dimethyl Sulfoxide (DMSO)	10%	78.1	72.0	%	Incubation: 100 mM Tris-HCl buffer, Assay: 100 mM Tris-HCl buffer, 0.2 MM NADH, 25 U/ml alcohol dehydrogenase from Saccharomyces cerevisiae	Incubation: 7, Assay: 7	Incubation: 50°C, Assay: 40°C	100 mM D-Erythrose, 9.1 mM D-Ribose 5-phosphate	Glyceraldehyde , Sedoheptulose-7-phosphate	0.5 mM MgCl₂, 0.1 mM ThDP	—	Water-incubated control (in %), assay in aqueous phase \| Uncertainty about whether assay really was in aqueous phase
Stability - Incubation	Activity after incubation (22 hours at 50°C), measured by absorbance spectrophotometry (NADH absorbance measurement, as reaction product glyceraldehyde reacts with alcohol dehydrogenase, 340 nm) in aqueous phase	Dimethyl Sulfoxide (DMSO)	10%	89.6	87.0	%	Incubation: 100 mM Tris-HCl buffer, Assay: 100 mM Tris-HCl buffer, 0.2 MM NADH, 25 U/ml alcohol dehydrogenase from Saccharomyces cerevisiae	Incubation: 7, Assay: 7	Incubation: 50°C, Assay: 40°C	100 mM D-Erythrose, 9.1 mM D-Ribose 5-phosphate	Glyceraldehyde , Sedoheptulose-7-phosphate	0.5 mM MgCl₂, 0.1 mM ThDP	—	Water-incubated control (in %), assay in aqueous phase \| Uncertainty about whether assay really was in aqueous phase
Stability - Incubation	Activity after incubation (6 hours at 50°C), measured by absorbance spectrophotometry (NADH absorbance measurement, as reaction product glyceraldehyde reacts with alcohol dehydrogenase, 340 nm) in aqueous phase	Dimethyl Sulfoxide (DMSO)	10%	95.9	98.5	%	Incubation: 100 mM Tris-HCl buffer, Assay: 100 mM Tris-HCl buffer, 0.2 MM NADH, 25 U/ml alcohol dehydrogenase from Saccharomyces cerevisiae	Incubation: 7, Assay: 7	Incubation: 50°C, Assay: 40°C	100 mM D-Erythrose, 9.1 mM D-Ribose 5-phosphate	Glyceraldehyde , Sedoheptulose-7-phosphate	0.5 mM MgCl₂, 0.1 mM ThDP	—	Water-incubated control (in %), assay in aqueous phase \| Uncertainty about whether assay really was in aqueous phase

Stability - Incubation

Activity after incubation (3 hours at 50°C), measured by absorbance spectrophotometry (NADH absorbance measurement, as reaction product glyceraldehyde reacts with alcohol dehydrogenase, 340 nm) in aqueous phase

Dimethyl Sulfoxide (DMSO)

50%

100.1

84.0

Incubation: 100 mM Tris-HCl buffer, Assay: 100 mM Tris-HCl buffer, 0.2 MM NADH, 25 U/ml alcohol dehydrogenase from Saccharomyces cerevisiae

Incubation: 7, Assay: 7

Incubation: 50°C, Assay: 40°C

100 mM D-Erythrose, 9.1 mM D-Ribose 5-phosphate

Glyceraldehyde, Sedoheptulose-7-phosphate

0.5 mM MgCl₂, 0.1 mM ThDP

—

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

Stability - Incubation

Activity after incubation (28 hours at 50°C), measured by absorbance spectrophotometry (NADH absorbance measurement, as reaction product glyceraldehyde reacts with alcohol dehydrogenase, 340 nm) in aqueous phase

Acetonitrile

50%

81.0

66.1

Incubation: 100 mM Tris-HCl buffer, Assay: 100 mM Tris-HCl buffer, 0.2 MM NADH, 25 U/ml alcohol dehydrogenase from Saccharomyces cerevisiae

Incubation: 7, Assay: 7

Incubation: 50°C, Assay: 40°C

100 mM D-Erythrose, 9.1 mM D-Ribose 5-phosphate

Glyceraldehyde, Sedoheptulose-7-phosphate

0.5 mM MgCl₂, 0.1 mM ThDP

—

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

Stability - Incubation

Activity after incubation (21 hours at 50°C), measured by absorbance spectrophotometry (NADH absorbance measurement, as reaction product glyceraldehyde reacts with alcohol dehydrogenase, 340 nm) in aqueous phase

Acetonitrile

50%

90.0

67.6

Incubation: 100 mM Tris-HCl buffer, Assay: 100 mM Tris-HCl buffer, 0.2 MM NADH, 25 U/ml alcohol dehydrogenase from Saccharomyces cerevisiae

Incubation: 7, Assay: 7

Incubation: 50°C, Assay: 40°C

100 mM D-Erythrose, 9.1 mM D-Ribose 5-phosphate

Glyceraldehyde, Sedoheptulose-7-phosphate

0.5 mM MgCl₂, 0.1 mM ThDP

—

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

Stability - Incubation

Activity after incubation (5 hours at 50°C), measured by absorbance spectrophotometry (NADH absorbance measurement, as reaction product glyceraldehyde reacts with alcohol dehydrogenase, 340 nm) in aqueous phase

Acetonitrile

50%

97.4

75.5

Incubation: 100 mM Tris-HCl buffer, Assay: 100 mM Tris-HCl buffer, 0.2 MM NADH, 25 U/ml alcohol dehydrogenase from Saccharomyces cerevisiae

Incubation: 7, Assay: 7

Incubation: 50°C, Assay: 40°C

100 mM D-Erythrose, 9.1 mM D-Ribose 5-phosphate

Glyceraldehyde, Sedoheptulose-7-phosphate

0.5 mM MgCl₂, 0.1 mM ThDP

—

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

Stability - Incubation

Acetonitrile

50%

100.1

69.3

Incubation: 100 mM Tris-HCl buffer, Assay: 100 mM Tris-HCl buffer, 0.2 MM NADH, 25 U/ml alcohol dehydrogenase from Saccharomyces cerevisiae

Incubation: 7, Assay: 7

Incubation: 50°C, Assay: 40°C

100 mM D-Erythrose, 9.1 mM D-Ribose 5-phosphate

Glyceraldehyde, Sedoheptulose-7-phosphate

0.5 mM MgCl₂, 0.1 mM ThDP

—

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

Stability - Incubation

Methanol

50%

81.0

75.3

Incubation: 100 mM Tris-HCl buffer, Assay: 100 mM Tris-HCl buffer, 0.2 MM NADH, 25 U/ml alcohol dehydrogenase from Saccharomyces cerevisiae

Incubation: 7, Assay: 7

Incubation: 50°C, Assay: 40°C

100 mM D-Erythrose, 9.1 mM D-Ribose 5-phosphate

Glyceraldehyde, Sedoheptulose-7-phosphate

0.5 mM MgCl₂, 0.1 mM ThDP

—

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

Stability - Incubation

Methanol

50%

90.0

77.3

Incubation: 100 mM Tris-HCl buffer, Assay: 100 mM Tris-HCl buffer, 0.2 MM NADH, 25 U/ml alcohol dehydrogenase from Saccharomyces cerevisiae

Incubation: 7, Assay: 7

Incubation: 50°C, Assay: 40°C

100 mM D-Erythrose, 9.1 mM D-Ribose 5-phosphate

Glyceraldehyde, Sedoheptulose-7-phosphate

0.5 mM MgCl₂, 0.1 mM ThDP

—

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

Stability - Incubation

Methanol

50%

97.4

77.8

Incubation: 100 mM Tris-HCl buffer, Assay: 100 mM Tris-HCl buffer, 0.2 MM NADH, 25 U/ml alcohol dehydrogenase from Saccharomyces cerevisiae

Incubation: 7, Assay: 7

Incubation: 50°C, Assay: 40°C

100 mM D-Erythrose, 9.1 mM D-Ribose 5-phosphate

Glyceraldehyde, Sedoheptulose-7-phosphate

0.5 mM MgCl₂, 0.1 mM ThDP

—

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

Stability - Incubation

Methanol

50%

100.1

72.0

Incubation: 100 mM Tris-HCl buffer, Assay: 100 mM Tris-HCl buffer, 0.2 MM NADH, 25 U/ml alcohol dehydrogenase from Saccharomyces cerevisiae

Incubation: 7, Assay: 7

Incubation: 50°C, Assay: 40°C

100 mM D-Erythrose, 9.1 mM D-Ribose 5-phosphate

Glyceraldehyde, Sedoheptulose-7-phosphate

0.5 mM MgCl₂, 0.1 mM ThDP

—

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

Stability - Incubation

Dimethyl Sulfoxide (DMSO)

50%

81.0

70.3

Incubation: 100 mM Tris-HCl buffer, Assay: 100 mM Tris-HCl buffer, 0.2 MM NADH, 25 U/ml alcohol dehydrogenase from Saccharomyces cerevisiae

Incubation: 7, Assay: 7

Incubation: 50°C, Assay: 40°C

100 mM D-Erythrose, 9.1 mM D-Ribose 5-phosphate

Glyceraldehyde, Sedoheptulose-7-phosphate

0.5 mM MgCl₂, 0.1 mM ThDP

—

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

Stability - Incubation

Dimethyl Sulfoxide (DMSO)

50%

90.0

68.3

Incubation: 100 mM Tris-HCl buffer, Assay: 100 mM Tris-HCl buffer, 0.2 MM NADH, 25 U/ml alcohol dehydrogenase from Saccharomyces cerevisiae

Incubation: 7, Assay: 7

Incubation: 50°C, Assay: 40°C

100 mM D-Erythrose, 9.1 mM D-Ribose 5-phosphate

Glyceraldehyde, Sedoheptulose-7-phosphate

0.5 mM MgCl₂, 0.1 mM ThDP

—

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

Stability - Incubation

Dimethyl Sulfoxide (DMSO)

50%

97.4

76.0

Incubation: 100 mM Tris-HCl buffer, Assay: 100 mM Tris-HCl buffer, 0.2 MM NADH, 25 U/ml alcohol dehydrogenase from Saccharomyces cerevisiae

Incubation: 7, Assay: 7

Incubation: 50°C, Assay: 40°C

100 mM D-Erythrose, 9.1 mM D-Ribose 5-phosphate

Glyceraldehyde, Sedoheptulose-7-phosphate

0.5 mM MgCl₂, 0.1 mM ThDP

—

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

Stability - Incubation

Activity after incubation (2 hours at 50°C), measured by absorbance spectrophotometry (NADH absorbance measurement, as reaction product glyceraldehyde reacts with alcohol dehydrogenase, 340 nm) in aqueous phase

Dimethyl Sulfoxide (DMSO)

10%

101.4

95.6

Incubation: 100 mM Tris-HCl buffer, Assay: 100 mM Tris-HCl buffer, 0.2 MM NADH, 25 U/ml alcohol dehydrogenase from Saccharomyces cerevisiae

Incubation: 7, Assay: 7

Incubation: 50°C, Assay: 40°C

100 mM D-Erythrose, 9.1 mM D-Ribose 5-phosphate

Glyceraldehyde, Sedoheptulose-7-phosphate

0.5 mM MgCl₂, 0.1 mM ThDP

—

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

Stability - Incubation

Activity after incubation (30 hours at 50°C), measured by absorbance spectrophotometry (NADH absorbance measurement, as reaction product glyceraldehyde reacts with alcohol dehydrogenase, 340 nm) in aqueous phase

Acetonitrile

10%

78.1

61.0

Incubation: 100 mM Tris-HCl buffer, Assay: 100 mM Tris-HCl buffer, 0.2 MM NADH, 25 U/ml alcohol dehydrogenase from Saccharomyces cerevisiae

Incubation: 7, Assay: 7

Incubation: 50°C, Assay: 40°C

100 mM D-Erythrose, 9.1 mM D-Ribose 5-phosphate

Glyceraldehyde, Sedoheptulose-7-phosphate

0.5 mM MgCl₂, 0.1 mM ThDP

—

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

Stability - Incubation

Activity after incubation (22 hours at 50°C), measured by absorbance spectrophotometry (NADH absorbance measurement, as reaction product glyceraldehyde reacts with alcohol dehydrogenase, 340 nm) in aqueous phase

Acetonitrile

10%

89.6

81.7

Incubation: 100 mM Tris-HCl buffer, Assay: 100 mM Tris-HCl buffer, 0.2 MM NADH, 25 U/ml alcohol dehydrogenase from Saccharomyces cerevisiae

Incubation: 7, Assay: 7

Incubation: 50°C, Assay: 40°C

100 mM D-Erythrose, 9.1 mM D-Ribose 5-phosphate

Glyceraldehyde, Sedoheptulose-7-phosphate

0.5 mM MgCl₂, 0.1 mM ThDP

—

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

Stability - Incubation

Activity after incubation (6 hours at 50°C), measured by absorbance spectrophotometry (NADH absorbance measurement, as reaction product glyceraldehyde reacts with alcohol dehydrogenase, 340 nm) in aqueous phase

Acetonitrile

10%

95.9

100.3

Incubation: 100 mM Tris-HCl buffer, Assay: 100 mM Tris-HCl buffer, 0.2 MM NADH, 25 U/ml alcohol dehydrogenase from Saccharomyces cerevisiae

Incubation: 7, Assay: 7

Incubation: 50°C, Assay: 40°C

100 mM D-Erythrose, 9.1 mM D-Ribose 5-phosphate

Glyceraldehyde, Sedoheptulose-7-phosphate

0.5 mM MgCl₂, 0.1 mM ThDP

—

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

Stability - Incubation

Acetonitrile

10%

101.4

100.9

Incubation: 100 mM Tris-HCl buffer, Assay: 100 mM Tris-HCl buffer, 0.2 MM NADH, 25 U/ml alcohol dehydrogenase from Saccharomyces cerevisiae

Incubation: 7, Assay: 7

Incubation: 50°C, Assay: 40°C

100 mM D-Erythrose, 9.1 mM D-Ribose 5-phosphate

Glyceraldehyde, Sedoheptulose-7-phosphate

0.5 mM MgCl₂, 0.1 mM ThDP

—

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

Stability - Incubation

Methanol

10%

78.1

67.1

Incubation: 100 mM Tris-HCl buffer, Assay: 100 mM Tris-HCl buffer, 0.2 MM NADH, 25 U/ml alcohol dehydrogenase from Saccharomyces cerevisiae

Incubation: 7, Assay: 7

Incubation: 50°C, Assay: 40°C

100 mM D-Erythrose, 9.1 mM D-Ribose 5-phosphate

Glyceraldehyde, Sedoheptulose-7-phosphate

0.5 mM MgCl₂, 0.1 mM ThDP

—

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

Stability - Incubation

Methanol

10%

89.6

79.1

Incubation: 100 mM Tris-HCl buffer, Assay: 100 mM Tris-HCl buffer, 0.2 MM NADH, 25 U/ml alcohol dehydrogenase from Saccharomyces cerevisiae

Incubation: 7, Assay: 7

Incubation: 50°C, Assay: 40°C

100 mM D-Erythrose, 9.1 mM D-Ribose 5-phosphate

Glyceraldehyde, Sedoheptulose-7-phosphate

0.5 mM MgCl₂, 0.1 mM ThDP

—

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

Stability - Incubation

Methanol

10%

95.9

105.6

Incubation: 100 mM Tris-HCl buffer, Assay: 100 mM Tris-HCl buffer, 0.2 MM NADH, 25 U/ml alcohol dehydrogenase from Saccharomyces cerevisiae

Incubation: 7, Assay: 7

Incubation: 50°C, Assay: 40°C

100 mM D-Erythrose, 9.1 mM D-Ribose 5-phosphate

Glyceraldehyde, Sedoheptulose-7-phosphate

0.5 mM MgCl₂, 0.1 mM ThDP

—

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

Stability - Incubation

Methanol

10%

101.4

110.3

Incubation: 100 mM Tris-HCl buffer, Assay: 100 mM Tris-HCl buffer, 0.2 MM NADH, 25 U/ml alcohol dehydrogenase from Saccharomyces cerevisiae

Incubation: 7, Assay: 7

Incubation: 50°C, Assay: 40°C

100 mM D-Erythrose, 9.1 mM D-Ribose 5-phosphate

Glyceraldehyde, Sedoheptulose-7-phosphate

0.5 mM MgCl₂, 0.1 mM ThDP

—

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

Stability - Incubation

Dimethyl Sulfoxide (DMSO)

10%

78.1

72.0

Incubation: 100 mM Tris-HCl buffer, Assay: 100 mM Tris-HCl buffer, 0.2 MM NADH, 25 U/ml alcohol dehydrogenase from Saccharomyces cerevisiae

Incubation: 7, Assay: 7

Incubation: 50°C, Assay: 40°C

100 mM D-Erythrose, 9.1 mM D-Ribose 5-phosphate

Glyceraldehyde, Sedoheptulose-7-phosphate

0.5 mM MgCl₂, 0.1 mM ThDP

—

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

Stability - Incubation

Dimethyl Sulfoxide (DMSO)

10%

89.6

87.0

Incubation: 100 mM Tris-HCl buffer, Assay: 100 mM Tris-HCl buffer, 0.2 MM NADH, 25 U/ml alcohol dehydrogenase from Saccharomyces cerevisiae

Incubation: 7, Assay: 7

Incubation: 50°C, Assay: 40°C

100 mM D-Erythrose, 9.1 mM D-Ribose 5-phosphate

Glyceraldehyde, Sedoheptulose-7-phosphate

0.5 mM MgCl₂, 0.1 mM ThDP

—

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

Stability - Incubation

Dimethyl Sulfoxide (DMSO)

10%

95.9

98.5

Incubation: 100 mM Tris-HCl buffer, Assay: 100 mM Tris-HCl buffer, 0.2 MM NADH, 25 U/ml alcohol dehydrogenase from Saccharomyces cerevisiae

Incubation: 7, Assay: 7

Incubation: 50°C, Assay: 40°C

100 mM D-Erythrose, 9.1 mM D-Ribose 5-phosphate

Glyceraldehyde, Sedoheptulose-7-phosphate

0.5 mM MgCl₂, 0.1 mM ThDP

—

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

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Visualization : Stability — Incubation

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Structure

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Transketolase (gene TKtmar) from Thermotoga maritima DSM3109

Enzyme Description

Sequence

Experimental Data (24 measurements)

Visualization : Stability — Incubation

Structure

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