"Split-gene" reconstituted transketolase (gene ChTK-F) from Carboxydothermus hydrogenoformans

Enzyme Description

Species
Carboxydothermus hydrogenoformans β†—
Extremophile
Yes "hyperthermophilic" organism
EC Number
2.2.1.1 β†—

Sequence

Length: 592 amino acids
MQDEILNLKLIANQLRQHVVKMVGEANSGHPGGSLSAADILAVLFFKEMRIDPANPKWQDRDRFVLSKGHASPVLYAALAERGFFPKEWLSQFRKINSPLQGHPDMKKVPGVEMSTGSLGQGFSTAVGMALGLKLDRSPARVYVLLGDGEIQEGIVWEAAMAAAHYKLNNLTAILDYNGLQIDGPVQEVMNPEPVADKWRSFGFKVITVDGHNIPEIINAIDAARLHLEGPTIIIAKTVKGKGVSFMENRVEWHGSAPKPEQVAEALSELQVGREKLWEEMGGIATREAYGKALVELGQENPKIVVLDADLSKSTKTSDFAKAFPERFFNMGIAEQNLMGVAAGLSTVGKIPFASTFAVFAAGRAFEIIRNSICYPKLNVKIAATHAGLTVGEDGASHQAIEDLALMRVLPNMQVFVPADAAQTRAIVKKAAEIEGPVYIRLGRSGVPEVFSPDIRFEPGRGTVLKEGKDVTIVALGIMTAKALEAAKMLEAEGIAARVVDMASLKPIDRELLVESARLTGAVVTAEEHSVIGGLGSAVAEVLSEEYPIPVVKVGVNDVFGESGTPQALLEKYGLTARDVVAAVQKALTLKR
Paul James et al. (2020) β€” A 'Split-Gene' Transketolase From the Hyper-Thermophilic Bacterium Carboxydothermus hydrogenoformans: Structure and Biochemical Characterization
Frontiers in Microbiology  Β· doi:10.3389/fmicb.2020.592353 β†—  Β· Stability - Incubation
18 measurements ▼
Database ID
β€”
Sequence Annotation
Explicit - Provided
Protein Source
Recombinant, host bacterium Escherichia coli BL21 (DE3)

Experimental Data (18 measurements)

18 measurements
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 (1 hour at room temperature), measured by HPLC (erythrulose production and Hydroxypyruvic acid consumption) in aqueous phase Dimethyl Sulfoxide (DMSO) 10% 100.0 47.9 % Incubation: 25 mM Tris-HCl buffer, 100 mM NaCl, Assay: 10 mM HEPES buffer, 100 mM NaCl, 9 mM CaCl2 Incubation: 7.5, Assay: 7.2 Incubation: Room temperature, Assay: Room temperature ? mM Glycoaldehyde, ? mM Hydroxypyruvic acid Carbon Dioxide (CO2) , Erythrulose Assay: 2 mM thiamine pyrophosphate β€” Non-incubated control (in %), assay in aqueous phase | Diluted organic solvent in asssay ("no greated than 7,5%")
Stability - Incubation Activity after incubation (1 hour at room temperature), measured by HPLC (erythrulose production and Hydroxypyruvic acid consumption) in aqueous phase Acetonitrile 50% 100.0 49.9 % Incubation: 25 mM Tris-HCl buffer, 100 mM NaCl, Assay: 10 mM HEPES buffer, 100 mM NaCl, 9 mM CaCl2 Incubation: 7.5, Assay: 7.2 Incubation: Room temperature, Assay: Room temperature ? mM Glycoaldehyde, ? mM Hydroxypyruvic acid Carbon Dioxide (CO2) , Erythrulose Assay: 2 mM thiamine pyrophosphate β€” Non-incubated control (in %), assay in aqueous phase | Diluted organic solvent in asssay ("no greated than 7,5%")
Stability - Incubation Activity after incubation (1 hour at room temperature), measured by HPLC (erythrulose production and Hydroxypyruvic acid consumption) in aqueous phase Acetonitrile 25% 100.0 51.9 % Incubation: 25 mM Tris-HCl buffer, 100 mM NaCl, Assay: 10 mM HEPES buffer, 100 mM NaCl, 9 mM CaCl2 Incubation: 7.5, Assay: 7.2 Incubation: Room temperature, Assay: Room temperature ? mM Glycoaldehyde, ? mM Hydroxypyruvic acid Carbon Dioxide (CO2) , Erythrulose Assay: 2 mM thiamine pyrophosphate β€” Non-incubated control (in %), assay in aqueous phase | Diluted organic solvent in asssay ("no greated than 7,5%")
Stability - Incubation Activity after incubation (1 hour at room temperature), measured by HPLC (erythrulose production and Hydroxypyruvic acid consumption) in aqueous phase Acetonitrile 10% 100.0 52.1 % Incubation: 25 mM Tris-HCl buffer, 100 mM NaCl, Assay: 10 mM HEPES buffer, 100 mM NaCl, 9 mM CaCl2 Incubation: 7.5, Assay: 7.2 Incubation: Room temperature, Assay: Room temperature ? mM Glycoaldehyde, ? mM Hydroxypyruvic acid Carbon Dioxide (CO2) , Erythrulose Assay: 2 mM thiamine pyrophosphate β€” Non-incubated control (in %), assay in aqueous phase | Diluted organic solvent in asssay ("no greated than 7,5%")
Stability - Incubation Activity after incubation (1 hour at room temperature), measured by HPLC (erythrulose production and Hydroxypyruvic acid consumption) in aqueous phase Acetone 50% 100.0 52.1 % Incubation: 25 mM Tris-HCl buffer, 100 mM NaCl, Assay: 10 mM HEPES buffer, 100 mM NaCl, 9 mM CaCl2 Incubation: 7.5, Assay: 7.2 Incubation: Room temperature, Assay: Room temperature ? mM Glycoaldehyde, ? mM Hydroxypyruvic acid Carbon Dioxide (CO2) , Erythrulose Assay: 2 mM thiamine pyrophosphate β€” Non-incubated control (in %), assay in aqueous phase | Diluted organic solvent in asssay ("no greated than 7,5%")
Stability - Incubation Activity after incubation (1 hour at room temperature), measured by HPLC (erythrulose production and Hydroxypyruvic acid consumption) in aqueous phase Acetone 25% 100.0 49.9 % Incubation: 25 mM Tris-HCl buffer, 100 mM NaCl, Assay: 10 mM HEPES buffer, 100 mM NaCl, 9 mM CaCl2 Incubation: 7.5, Assay: 7.2 Incubation: Room temperature, Assay: Room temperature ? mM Glycoaldehyde, ? mM Hydroxypyruvic acid Carbon Dioxide (CO2) , Erythrulose Assay: 2 mM thiamine pyrophosphate β€” Non-incubated control (in %), assay in aqueous phase | Diluted organic solvent in asssay ("no greated than 7,5%")
Stability - Incubation Activity after incubation (1 hour at room temperature), measured by HPLC (erythrulose production and Hydroxypyruvic acid consumption) in aqueous phase Acetone 10% 100.0 52.1 % Incubation: 25 mM Tris-HCl buffer, 100 mM NaCl, Assay: 10 mM HEPES buffer, 100 mM NaCl, 9 mM CaCl2 Incubation: 7.5, Assay: 7.2 Incubation: Room temperature, Assay: Room temperature ? mM Glycoaldehyde, ? mM Hydroxypyruvic acid Carbon Dioxide (CO2) , Erythrulose Assay: 2 mM thiamine pyrophosphate β€” Non-incubated control (in %), assay in aqueous phase | Diluted organic solvent in asssay ("no greated than 7,5%")
Stability - Incubation Activity after incubation (1 hour at room temperature), measured by HPLC (erythrulose production and Hydroxypyruvic acid consumption) in aqueous phase Dimethyl Sulfoxide (DMSO) 50% 100.0 51.9 % Incubation: 25 mM Tris-HCl buffer, 100 mM NaCl, Assay: 10 mM HEPES buffer, 100 mM NaCl, 9 mM CaCl2 Incubation: 7.5, Assay: 7.2 Incubation: Room temperature, Assay: Room temperature ? mM Glycoaldehyde, ? mM Hydroxypyruvic acid Carbon Dioxide (CO2) , Erythrulose Assay: 2 mM thiamine pyrophosphate β€” Non-incubated control (in %), assay in aqueous phase | Diluted organic solvent in asssay ("no greated than 7,5%")
Stability - Incubation Activity after incubation (1 hour at room temperature), measured by HPLC (erythrulose production and Hydroxypyruvic acid consumption) in aqueous phase Dimethyl Sulfoxide (DMSO) 25% 100.0 49.9 % Incubation: 25 mM Tris-HCl buffer, 100 mM NaCl, Assay: 10 mM HEPES buffer, 100 mM NaCl, 9 mM CaCl2 Incubation: 7.5, Assay: 7.2 Incubation: Room temperature, Assay: Room temperature ? mM Glycoaldehyde, ? mM Hydroxypyruvic acid Carbon Dioxide (CO2) , Erythrulose Assay: 2 mM thiamine pyrophosphate β€” Non-incubated control (in %), assay in aqueous phase | Diluted organic solvent in asssay ("no greated than 7,5%")
Stability - Incubation Activity after incubation (1 hour at room temperature), measured by HPLC (erythrulose production and Hydroxypyruvic acid consumption) in aqueous phase Methanol 10% 100.0 105.0 % Incubation: 25 mM Tris-HCl buffer, 100 mM NaCl, Assay: 10 mM HEPES buffer, 100 mM NaCl, 9 mM CaCl2 Incubation: 7.5, Assay: 7.2 Incubation: Room temperature, Assay: Room temperature ? mM Glycoaldehyde, ? mM Hydroxypyruvic acid Carbon Dioxide (CO2) , Erythrulose Assay: 2 mM thiamine pyrophosphate β€” Non-incubated control (in %), assay in aqueous phase | Diluted organic solvent in asssay ("no greated than 7,5%")
Stability - Incubation Activity after incubation (1 hour at room temperature), measured by HPLC (erythrulose production and Hydroxypyruvic acid consumption) in aqueous phase Isopropanol 50% 100.0 60.0 % Incubation: 25 mM Tris-HCl buffer, 100 mM NaCl, Assay: 10 mM HEPES buffer, 100 mM NaCl, 9 mM CaCl2 Incubation: 7.5, Assay: 7.2 Incubation: Room temperature, Assay: Room temperature ? mM Glycoaldehyde, ? mM Hydroxypyruvic acid Carbon Dioxide (CO2) , Erythrulose Assay: 2 mM thiamine pyrophosphate β€” Non-incubated control (in %), assay in aqueous phase | Diluted organic solvent in asssay ("no greated than 7,5%")
Stability - Incubation Activity after incubation (1 hour at room temperature), measured by HPLC (erythrulose production and Hydroxypyruvic acid consumption) in aqueous phase Isopropanol 25% 100.0 58.0 % Incubation: 25 mM Tris-HCl buffer, 100 mM NaCl, Assay: 10 mM HEPES buffer, 100 mM NaCl, 9 mM CaCl2 Incubation: 7.5, Assay: 7.2 Incubation: Room temperature, Assay: Room temperature ? mM Glycoaldehyde, ? mM Hydroxypyruvic acid Carbon Dioxide (CO2) , Erythrulose Assay: 2 mM thiamine pyrophosphate β€” Non-incubated control (in %), assay in aqueous phase | Diluted organic solvent in asssay ("no greated than 7,5%")
Stability - Incubation Activity after incubation (1 hour at room temperature), measured by HPLC (erythrulose production and Hydroxypyruvic acid consumption) in aqueous phase Isopropanol 10% 100.0 60.0 % Incubation: 25 mM Tris-HCl buffer, 100 mM NaCl, Assay: 10 mM HEPES buffer, 100 mM NaCl, 9 mM CaCl2 Incubation: 7.5, Assay: 7.2 Incubation: Room temperature, Assay: Room temperature ? mM Glycoaldehyde, ? mM Hydroxypyruvic acid Carbon Dioxide (CO2) , Erythrulose Assay: 2 mM thiamine pyrophosphate β€” Non-incubated control (in %), assay in aqueous phase | Diluted organic solvent in asssay ("no greated than 7,5%")
Stability - Incubation Activity after incubation (1 hour at room temperature), measured by HPLC (erythrulose production and Hydroxypyruvic acid consumption) in aqueous phase Ethanol 50% 100.0 44.0 % Incubation: 25 mM Tris-HCl buffer, 100 mM NaCl, Assay: 10 mM HEPES buffer, 100 mM NaCl, 9 mM CaCl2 Incubation: 7.5, Assay: 7.2 Incubation: Room temperature, Assay: Room temperature ? mM Glycoaldehyde, ? mM Hydroxypyruvic acid Carbon Dioxide (CO2) , Erythrulose Assay: 2 mM thiamine pyrophosphate β€” Non-incubated control (in %), assay in aqueous phase | Diluted organic solvent in asssay ("no greated than 7,5%")
Stability - Incubation Activity after incubation (1 hour at room temperature), measured by HPLC (erythrulose production and Hydroxypyruvic acid consumption) in aqueous phase Ethanol 25% 100.0 42.0 % Incubation: 25 mM Tris-HCl buffer, 100 mM NaCl, Assay: 10 mM HEPES buffer, 100 mM NaCl, 9 mM CaCl2 Incubation: 7.5, Assay: 7.2 Incubation: Room temperature, Assay: Room temperature ? mM Glycoaldehyde, ? mM Hydroxypyruvic acid Carbon Dioxide (CO2) , Erythrulose Assay: 2 mM thiamine pyrophosphate β€” Non-incubated control (in %), assay in aqueous phase | Diluted organic solvent in asssay ("no greated than 7,5%")
Stability - Incubation Activity after incubation (1 hour at room temperature), measured by HPLC (erythrulose production and Hydroxypyruvic acid consumption) in aqueous phase Ethanol 10% 100.0 70.1 % Incubation: 25 mM Tris-HCl buffer, 100 mM NaCl, Assay: 10 mM HEPES buffer, 100 mM NaCl, 9 mM CaCl2 Incubation: 7.5, Assay: 7.2 Incubation: Room temperature, Assay: Room temperature ? mM Glycoaldehyde, ? mM Hydroxypyruvic acid Carbon Dioxide (CO2) , Erythrulose Assay: 2 mM thiamine pyrophosphate β€” Non-incubated control (in %), assay in aqueous phase | Diluted organic solvent in asssay ("no greated than 7,5%")
Stability - Incubation Activity after incubation (1 hour at room temperature), measured by HPLC (erythrulose production and Hydroxypyruvic acid consumption) in aqueous phase Methanol 50% 100.0 40.0 % Incubation: 25 mM Tris-HCl buffer, 100 mM NaCl, Assay: 10 mM HEPES buffer, 100 mM NaCl, 9 mM CaCl2 Incubation: 7.5, Assay: 7.2 Incubation: Room temperature, Assay: Room temperature ? mM Glycoaldehyde, ? mM Hydroxypyruvic acid Carbon Dioxide (CO2) , Erythrulose Assay: 2 mM thiamine pyrophosphate β€” Non-incubated control (in %), assay in aqueous phase | Diluted organic solvent in asssay ("no greated than 7,5%")
Stability - Incubation Activity after incubation (1 hour at room temperature), measured by HPLC (erythrulose production and Hydroxypyruvic acid consumption) in aqueous phase Methanol 25% 100.0 54.9 % Incubation: 25 mM Tris-HCl buffer, 100 mM NaCl, Assay: 10 mM HEPES buffer, 100 mM NaCl, 9 mM CaCl2 Incubation: 7.5, Assay: 7.2 Incubation: Room temperature, Assay: Room temperature ? mM Glycoaldehyde, ? mM Hydroxypyruvic acid Carbon Dioxide (CO2) , Erythrulose Assay: 2 mM thiamine pyrophosphate β€” Non-incubated control (in %), assay in aqueous phase | Diluted organic solvent in asssay ("no greated than 7,5%")

Visualization : Stability β€” Incubation

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

Structure

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