Transketolase (gene TKtmar) from Thermotoga maritima DSM3109
Enzyme Description
Species
Extremophile
Yes
"hyperthermophilic" organism
EC Number
Sequence
MERFPYEKLPESELKELKELGRLCRGDILKMTYIANSGHPGGSMSSIDLYLTVFKYAKLRPVDDPARDRIVISHGHTSPGVYAAMARLGFVDLDEVLAGFRHPASVFEGHVTRGVGIIDWTTGNLGQGLSAGLGFALASRFTGKDYHVFVLMSDAEQAKGQVAEARRVAKKYGVTNLTVIIDYNDAQISGRARDVMPVNIKENYLADGWRVIEIDGHDYEQIYLALKEAVEDELNPVAIIAKTVMGKGVSFMENEVKYHGKPLNREELEKALAELGIENDVDVYIEKRKQLPVEKHKKVYKTYPIKIDTGEPITYTSPTDNRSAFGKAILDLVKKNVNNPETTPIVAVDCDLKGSVKLDLLDKEFPERLLEVGVQEHNAAAMAGALSAEGVITFFADFGVFGISETYNQHRLNAINGTNLKVVVTHCGLNVGEDGKTHHGLDYVSGPMNWYGFKVIVPGDPNQTDRVVRYAAKEYGNFVIAMGRSKLPIILDENGKPFFGEGYTFEYGKIDVVRKGDDAVIITYGSTLCEAVNAADELKKEGVNVAVLNVSCPVDLDIETLKMVDGKPVLVVEDHNVFTGLGSFLGTTLLENGIIPKKYVRVGVPEFAVSGSYTMLYKLYGLDKDGIISRLREML
Max CΓ‘rdenas-FernΓ‘ndez et al. (2021)
β Characterisation of a hyperthermophilic transketolase from Thermotoga maritima DSM3109 as a biocatalyst for 7-keto-octuronic acid synthesis
Organic & Biomolecular Chemistry
Β· doi:10.1039/d1ob01237a β
Β· Stability - Incubation
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Database ID
UniParc: UPI00000D3857 β
Sequence Annotation
Explicit - Provided GenBank Accession Number
Protein Source
Recombinant, host bacterium Escherichia coli BL21 (DE3)
Experimental Data (24 measurements)
24 measurements β page 1 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 (3 hours at 50Β°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: 50Β°C, Assay: 40Β°C | 100 mM D-Erythrose, 9.1 mM D-Ribose 5-phosphate | Glyceraldehyde , Sedoheptulose-7-phosphate | 0.5 mM MgClβ, 0.1 mM ThDP | β | 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 50Β°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: 50Β°C, Assay: 40Β°C | 100 mM D-Erythrose, 9.1 mM D-Ribose 5-phosphate | Glyceraldehyde , Sedoheptulose-7-phosphate | 0.5 mM MgClβ, 0.1 mM ThDP | β | 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 50Β°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: 50Β°C, Assay: 40Β°C | 100 mM D-Erythrose, 9.1 mM D-Ribose 5-phosphate | Glyceraldehyde , Sedoheptulose-7-phosphate | 0.5 mM MgClβ, 0.1 mM ThDP | β | 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 50Β°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: 50Β°C, Assay: 40Β°C | 100 mM D-Erythrose, 9.1 mM D-Ribose 5-phosphate | Glyceraldehyde , Sedoheptulose-7-phosphate | 0.5 mM MgClβ, 0.1 mM ThDP | β | 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 50Β°C), measured by absorbance spectrophotometry (NADH absorbance measurement, as reaction product glyceraldehyde reacts with alcohol dehydrogenase, 340 nm) in aqueous phase | 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: 50Β°C, Assay: 40Β°C | 100 mM D-Erythrose, 9.1 mM D-Ribose 5-phosphate | Glyceraldehyde , Sedoheptulose-7-phosphate | 0.5 mM MgClβ, 0.1 mM ThDP | β | 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 50Β°C), measured by absorbance spectrophotometry (NADH absorbance measurement, as reaction product glyceraldehyde reacts with alcohol dehydrogenase, 340 nm) in aqueous phase | 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: 50Β°C, Assay: 40Β°C | 100 mM D-Erythrose, 9.1 mM D-Ribose 5-phosphate | Glyceraldehyde , Sedoheptulose-7-phosphate | 0.5 mM MgClβ, 0.1 mM ThDP | β | 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 50Β°C), measured by absorbance spectrophotometry (NADH absorbance measurement, as reaction product glyceraldehyde reacts with alcohol dehydrogenase, 340 nm) in aqueous phase | 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: 50Β°C, Assay: 40Β°C | 100 mM D-Erythrose, 9.1 mM D-Ribose 5-phosphate | Glyceraldehyde , Sedoheptulose-7-phosphate | 0.5 mM MgClβ, 0.1 mM ThDP | β | 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 50Β°C), measured by absorbance spectrophotometry (NADH absorbance measurement, as reaction product glyceraldehyde reacts with alcohol dehydrogenase, 340 nm) in aqueous phase | 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: 50Β°C, Assay: 40Β°C | 100 mM D-Erythrose, 9.1 mM D-Ribose 5-phosphate | Glyceraldehyde , Sedoheptulose-7-phosphate | 0.5 mM MgClβ, 0.1 mM ThDP | β | 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 50Β°C), measured by absorbance spectrophotometry (NADH absorbance measurement, as reaction product glyceraldehyde reacts with alcohol dehydrogenase, 340 nm) in aqueous phase | 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: 50Β°C, Assay: 40Β°C | 100 mM D-Erythrose, 9.1 mM D-Ribose 5-phosphate | Glyceraldehyde , Sedoheptulose-7-phosphate | 0.5 mM MgClβ, 0.1 mM ThDP | β | 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 50Β°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% | 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: 50Β°C, Assay: 40Β°C | 100 mM D-Erythrose, 9.1 mM D-Ribose 5-phosphate | Glyceraldehyde , Sedoheptulose-7-phosphate | 0.5 mM MgClβ, 0.1 mM ThDP | β | 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 50Β°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% | 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: 50Β°C, Assay: 40Β°C | 100 mM D-Erythrose, 9.1 mM D-Ribose 5-phosphate | Glyceraldehyde , Sedoheptulose-7-phosphate | 0.5 mM MgClβ, 0.1 mM ThDP | β | 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 50Β°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% | 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: 50Β°C, Assay: 40Β°C | 100 mM D-Erythrose, 9.1 mM D-Ribose 5-phosphate | Glyceraldehyde , Sedoheptulose-7-phosphate | 0.5 mM MgClβ, 0.1 mM ThDP | β | 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 50Β°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: 50Β°C, Assay: 40Β°C | 100 mM D-Erythrose, 9.1 mM D-Ribose 5-phosphate | Glyceraldehyde , Sedoheptulose-7-phosphate | 0.5 mM MgClβ, 0.1 mM ThDP | β | 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 50Β°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: 50Β°C, Assay: 40Β°C | 100 mM D-Erythrose, 9.1 mM D-Ribose 5-phosphate | Glyceraldehyde , Sedoheptulose-7-phosphate | 0.5 mM MgClβ, 0.1 mM ThDP | β | 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 50Β°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: 50Β°C, Assay: 40Β°C | 100 mM D-Erythrose, 9.1 mM D-Ribose 5-phosphate | Glyceraldehyde , Sedoheptulose-7-phosphate | 0.5 mM MgClβ, 0.1 mM ThDP | β | 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 50Β°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: 50Β°C, Assay: 40Β°C | 100 mM D-Erythrose, 9.1 mM D-Ribose 5-phosphate | Glyceraldehyde , Sedoheptulose-7-phosphate | 0.5 mM MgClβ, 0.1 mM ThDP | β | 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 50Β°C), measured by absorbance spectrophotometry (NADH absorbance measurement, as reaction product glyceraldehyde reacts with alcohol dehydrogenase, 340 nm) in aqueous phase | 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: 50Β°C, Assay: 40Β°C | 100 mM D-Erythrose, 9.1 mM D-Ribose 5-phosphate | Glyceraldehyde , Sedoheptulose-7-phosphate | 0.5 mM MgClβ, 0.1 mM ThDP | β | 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 50Β°C), measured by absorbance spectrophotometry (NADH absorbance measurement, as reaction product glyceraldehyde reacts with alcohol dehydrogenase, 340 nm) in aqueous phase | 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: 50Β°C, Assay: 40Β°C | 100 mM D-Erythrose, 9.1 mM D-Ribose 5-phosphate | Glyceraldehyde , Sedoheptulose-7-phosphate | 0.5 mM MgClβ, 0.1 mM ThDP | β | 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 50Β°C), measured by absorbance spectrophotometry (NADH absorbance measurement, as reaction product glyceraldehyde reacts with alcohol dehydrogenase, 340 nm) in aqueous phase | 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: 50Β°C, Assay: 40Β°C | 100 mM D-Erythrose, 9.1 mM D-Ribose 5-phosphate | Glyceraldehyde , Sedoheptulose-7-phosphate | 0.5 mM MgClβ, 0.1 mM ThDP | β | 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 50Β°C), measured by absorbance spectrophotometry (NADH absorbance measurement, as reaction product glyceraldehyde reacts with alcohol dehydrogenase, 340 nm) in aqueous phase | 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: 50Β°C, Assay: 40Β°C | 100 mM D-Erythrose, 9.1 mM D-Ribose 5-phosphate | Glyceraldehyde , Sedoheptulose-7-phosphate | 0.5 mM MgClβ, 0.1 mM ThDP | β | Water-incubated control (in %), assay in aqueous phase | Uncertainty about whether assay really was in aqueous phase |
Visualization : Stability β Incubation
One bar per measurement. Colour = solvent, shade = solvent volume.