Cloned (Comment) | Organism |
---|---|
gene gldA, DNA and amino acid sequence determination and analysis, sequence comparisons and phylogenetic analysis, recombinant expression of His-tagged enzyme in Escherichia coli strain BL21(DE3) | Thermoanaerobacterium thermosaccharolyticum |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
Ca2+ | - |
Thermoanaerobacterium thermosaccharolyticum | |
SDS | - |
Thermoanaerobacterium thermosaccharolyticum | |
Tween 80 | - |
Thermoanaerobacterium thermosaccharolyticum | |
Zn2+ | inhibits 24% at 1 mM | Thermoanaerobacterium thermosaccharolyticum |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.06 | - |
NADH | pH 6.0, 60°C, recombinant enzyme | Thermoanaerobacterium thermosaccharolyticum | |
1.08 | - |
Glycerone | pH 6.0, 60°C, recombinant enzyme | Thermoanaerobacterium thermosaccharolyticum | |
30.29 | - |
glycerol | pH 8.0, 60°C, recombinant enzyme | Thermoanaerobacterium thermosaccharolyticum |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
Co2+ | presence of Mn2+ enhances the enzyme activity by 31.4% | Thermoanaerobacterium thermosaccharolyticum | |
Cu2+ | presence of Mn2+ enhances the enzyme activity by 37.0% | Thermoanaerobacterium thermosaccharolyticum | |
Mn2+ | presence of Mn2+ enhances the enzyme activity by 79.5% | Thermoanaerobacterium thermosaccharolyticum | |
additional information | three highly conserved enzyme residues, Asp171, His254, and His271, are associated with metal ion binding | Thermoanaerobacterium thermosaccharolyticum |
Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
---|---|---|---|
40400 | - |
- |
Thermoanaerobacterium thermosaccharolyticum |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
glycerol + NAD+ | Thermoanaerobacterium thermosaccharolyticum | - |
glycerone + NADH + H+ | - |
r | |
glycerol + NAD+ | Thermoanaerobacterium thermosaccharolyticum DSM 571 | - |
glycerone + NADH + H+ | - |
r |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Thermoanaerobacterium thermosaccharolyticum | - |
- |
- |
Thermoanaerobacterium thermosaccharolyticum DSM 571 | - |
- |
- |
Purification (Comment) | Organism |
---|---|
recombinant His-tagged enzyme from Escherichia coli strain BL21(DE3) by nickel affinity chromatography | Thermoanaerobacterium thermosaccharolyticum |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
glycerol + NAD+ | - |
Thermoanaerobacterium thermosaccharolyticum | glycerone + NADH + H+ | - |
r | |
glycerol + NAD+ | glycerone reduction is the dominant reaction | Thermoanaerobacterium thermosaccharolyticum | glycerone + NADH + H+ | i.e. 1,3-dihydroxypropranone | r | |
glycerol + NAD+ | - |
Thermoanaerobacterium thermosaccharolyticum DSM 571 | glycerone + NADH + H+ | - |
r | |
glycerol + NAD+ | glycerone reduction is the dominant reaction | Thermoanaerobacterium thermosaccharolyticum DSM 571 | glycerone + NADH + H+ | i.e. 1,3-dihydroxypropranone | r | |
additional information | enzyme TtGlyDH preferentially catalyzes 1,3-dihydroxypropranone reduction rather than alcohol compound oxidation. Glycerol oxidization activity is faintly detected in the presence of a high concentration of glycerol (137 mM). No activity is detected with primary alcohols or diols. The highest glycerol oxidation activity is observed at the optimal growth temperature of 60°C in Tris-HCl buffer (50 mM, pH 8.0). Maximum DHA reduction activity is also observed at 60°C, and TtGlyDH exhibits the highest activity in an acetate buffer, compared with 91% maximum activity in an imidazole buffer at the same pH of 6.0 | Thermoanaerobacterium thermosaccharolyticum | ? | - |
? | |
additional information | enzyme TtGlyDH preferentially catalyzes 1,3-dihydroxypropranone reduction rather than alcohol compound oxidation. Glycerol oxidization activity is faintly detected in the presence of a high concentration of glycerol (137 mM). No activity is detected with primary alcohols or diols. The highest glycerol oxidation activity is observed at the optimal growth temperature of 60°C in Tris-HCl buffer (50 mM, pH 8.0). Maximum DHA reduction activity is also observed at 60°C, and TtGlyDH exhibits the highest activity in an acetate buffer, compared with 91% maximum activity in an imidazole buffer at the same pH of 6.0 | Thermoanaerobacterium thermosaccharolyticum DSM 571 | ? | - |
? |
Subunits | Comment | Organism |
---|---|---|
? | x * 40400, recombinant enzyme, SDS-PAGE | Thermoanaerobacterium thermosaccharolyticum |
Synonyms | Comment | Organism |
---|---|---|
GldA | - |
Thermoanaerobacterium thermosaccharolyticum |
GlyDH | - |
Thermoanaerobacterium thermosaccharolyticum |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
60 | - |
both reaction directions | Thermoanaerobacterium thermosaccharolyticum |
Temperature Stability Minimum [°C] | Temperature Stability Maximum [°C] | Comment | Organism |
---|---|---|---|
55 | - |
recombinant enzyme, maintaining 70% activity after 2 h | Thermoanaerobacterium thermosaccharolyticum |
60 | - |
recombinant enzyme, maintaining 65% activity after 2 h | Thermoanaerobacterium thermosaccharolyticum |
60 | 70 | recombinant enzyme, maintaining over 80% activity after 30 min | Thermoanaerobacterium thermosaccharolyticum |
70 | - |
recombinant enzyme, maintaining 19% activity after 2 h | Thermoanaerobacterium thermosaccharolyticum |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
1.89 | - |
glycerol | pH 8.0, 60°C, recombinant enzyme | Thermoanaerobacterium thermosaccharolyticum | |
98.44 | - |
Glycerone | pH 6.0, 60°C, recombinant enzyme | Thermoanaerobacterium thermosaccharolyticum |
pH Minimum | pH Maximum | Comment | Organism |
---|---|---|---|
6 | - |
glycerone reduction | Thermoanaerobacterium thermosaccharolyticum |
8 | - |
glycerol oxidation | Thermoanaerobacterium thermosaccharolyticum |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
additional information | the enzyme's atypical dinucleotide binding motif (GGG motif) and basic residue Arg43 are both related to dinucleotide binding | Thermoanaerobacterium thermosaccharolyticum | |
NAD+ | - |
Thermoanaerobacterium thermosaccharolyticum | |
NADH | - |
Thermoanaerobacterium thermosaccharolyticum |
General Information | Comment | Organism |
---|---|---|
evolution | the enzyme belongs to the Fe-ADH family. The GXGXXG motif is not present in TtGlyDH or other members of the Fe-ADH family, the GGG motif forms a more flexible turn and provides enough space to accommodate the pyrophosphate moiety of dinucleotides | Thermoanaerobacterium thermosaccharolyticum |
additional information | molecular modeling and site-directed mutagenesis analyses demonstrate that TtGlyDH has an atypical dinucleotide binding motif (GGG motif) and a basic residue Arg43, both related to dinucleotide binding | Thermoanaerobacterium thermosaccharolyticum |
physiological function | glycerol dehydrogenases (GlyDHs) are essential for glycerol metabolism in vivo, catalyzing its reversible reduction to 1,3-dihydroxypropranone | Thermoanaerobacterium thermosaccharolyticum |
kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.06 | - |
glycerol | pH 8.0, 60°C, recombinant enzyme | Thermoanaerobacterium thermosaccharolyticum | |
91.15 | - |
Glycerone | pH 6.0, 60°C, recombinant enzyme | Thermoanaerobacterium thermosaccharolyticum |