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Literature summary for 1.2.1.4 extracted from
- Molecular dynamics analysis of a rationally designed aldehyde dehydrogenase gives insights into improved activity for the non-native cofactor NAD (2020), ACS Synth. Biol., 9, 920-929 .
Application
| Application | Comment | Organism |
|---|---|---|
| synthesis | the aldehyde dehydrogenase from Thermoplasma acidophilum implemented as a key enzyme in a synthetic cellfree reaction cascade for the production of alcohols. Thermoplasma acidophilum enzyme TaALDH matches the cascade equirements regarding temperature stability, efficient heterologous expression in Escherichia coli, and exclusive activity for D-glyceraldehyde (not active on acetaldehyde). The remaining drawback is its cofactor preference toward NADP+ resulting in high KM for NAD+ and a rather low overall activity under cascade conditions | Thermoplasma acidophilum |
Protein Variants
| Protein Variants | Comment | Organism |
|---|---|---|
| D176S | site-directed mutagenesis, the mutation results in increased activity with NAD+ compared to NADP+ and the wild-type enzyme | Thermoplasma acidophilum |
| D176S/M262I | site-directed mutagenesis, the mutation results in increased activity with NAD+ compared to NADP+ and the wild-type enzyme | Thermoplasma acidophilum |
| D176S/S206K | site-directed mutagenesis, the mutation results in increased activity with NAD+ compared to NADP+ and the wild-type enzyme | Thermoplasma acidophilum |
| D176S/S206K/M262I | site-directed mutagenesis, the mutation results in increased activity with NAD+ compared to NADP+ and the wild-type enzyme | Thermoplasma acidophilum |
| D176S/S206K/M262I/W271Y/W275V | site-directed mutagenesis, the mutation results in increased activity with NAD+ compared to NADP+ and the wild-type enzyme | Thermoplasma acidophilum |
| F34M/Y399C/S405N | site-directed mutagenesis, crystal structure analysis | Thermoplasma acidophilum |
| M262I | site-directed mutagenesis, the mutation results in 30% higher activity with NAD+ compared to wild-type enzyme, the surface mutation M262I has influence on the solubility | Thermoplasma acidophilum |
| additional information | random mutagenesis approach for the aldehyde dehydrogenase of Thermoplasma acidophilum (TaALDH) to increase volumetric activity and slightly improve NAD+ acceptance. Roughly 450 mutants do not show any activity, and around 400 mutants have an activity below the template (M33) threshold. Saturation mutagenesis of the residues at the entrance of the substrate pocket can eliminate substrate inhibition. Molecular dynamics simulations show a significant gain of flexibility at the cofactor binding site for the final variant | Thermoplasma acidophilum |
| S175E | site-directed mutagenesis, the mutation results in increased activity with NAD+ compared to NADP+ and the wild-type enzyme | Thermoplasma acidophilum |
| S175E/D176S | site-directed mutagenesis, the mutation results in increased activity with NAD+ compared to NADP+ and the wild-type enzyme | Thermoplasma acidophilum |
| S175E/D176S/M262I | site-directed mutagenesis, the mutation results in increased activity with NAD+ compared to NADP+ and the wild-type enzyme | Thermoplasma acidophilum |
| S175E/D176S/S206K | site-directed mutagenesis, the mutation results in increased activity with NAD+ compared to NADP+ and the wild-type enzyme | Thermoplasma acidophilum |
| S175E/D176S/S206K/M262I | site-directed mutagenesis, the mutation results in increased activity with NAD+ compared to NADP+ and the wild-type enzyme | Thermoplasma acidophilum |
| S175E/M262I | site-directed mutagenesis, the mutation results in increased activity with NAD+ compared to NADP+ and the wild-type enzyme | Thermoplasma acidophilum |
| S175E/S206K | site-directed mutagenesis, the mutation results in increased activity with NAD+ compared to NADP+ and the wild-type enzyme | Thermoplasma acidophilum |
| S175E/S206K/M262I | site-directed mutagenesis, the mutation results in increased activity with NAD+ compared to NADP+ and the wild-type enzyme | Thermoplasma acidophilum |
| S206K | site-directed mutagenesis, the mutation results in increased activity with NAD+ compared to NADP+ and the wild-type enzyme | Thermoplasma acidophilum |
| S206K/M262I | site-directed mutagenesis, the mutation results in increased activity with NAD+ compared to NADP+ and the wild-type enzyme | Thermoplasma acidophilum |
Inhibitors
| Inhibitors | Comment | Organism | Structure |
|---|---|---|---|
| D-glyceraldehyde | substrate inhibition | Thermoplasma acidophilum |  |
Organism
| Organism | UniProt | Comment | Textmining |
|---|---|---|---|
| Thermoplasma acidophilum | Q9HL01 | - |
- |
| Thermoplasma acidophilum AMRC-C165 | Q9HL01 | - |
- |
| Thermoplasma acidophilum ATCC 25905 | Q9HL01 | - |
- |
| Thermoplasma acidophilum JCM 9062 | Q9HL01 | - |
- |
| Thermoplasma acidophilum NBRC 15155 | Q9HL01 | - |
- |
Substrates and Products (Substrate)
| Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| D-glyceraldehyde + NADP+ | - |
Thermoplasma acidophilum | D-glycerate + NADPH + H+ | - |
? | |
| D-glyceraldehyde + NADP+ | - |
Thermoplasma acidophilum JCM 9062 | D-glycerate + NADPH + H+ | - |
? | |
| D-glyceraldehyde + NADP+ | - |
Thermoplasma acidophilum AMRC-C165 | D-glycerate + NADPH + H+ | - |
? | |
| D-glyceraldehyde + NADP+ | - |
Thermoplasma acidophilum ATCC 25905 | D-glycerate + NADPH + H+ | - |
? | |
| D-glyceraldehyde + NADP+ | - |
Thermoplasma acidophilum NBRC 15155 | D-glycerate + NADPH + H+ | - |
? | |
| additional information | no activity with acetaldehyde | Thermoplasma acidophilum | ? | - |
- |
|
| additional information | no activity with acetaldehyde | Thermoplasma acidophilum JCM 9062 | ? | - |
- |
|
| additional information | no activity with acetaldehyde | Thermoplasma acidophilum AMRC-C165 | ? | - |
- |
|
| additional information | no activity with acetaldehyde | Thermoplasma acidophilum ATCC 25905 | ? | - |
- |
|
| additional information | no activity with acetaldehyde | Thermoplasma acidophilum NBRC 15155 | ? | - |
- |
|
Synonyms
| Synonyms | Comment | Organism |
|---|---|---|
| ALDH | - |
Thermoplasma acidophilum |
| Ta0439 | - |
Thermoplasma acidophilum |
| TaAlDH | - |
Thermoplasma acidophilum |
Temperature Stability [°C]
| Temperature Stability Minimum [°C] | Temperature Stability Maximum [°C] | Comment | Organism |
|---|---|---|---|
| 7 | 9.5 | Tm of wild-type enzyme | Thermoplasma acidophilum |
| 72.5 | - |
Tm of mutant D176S/S206K/M262I/W271Y/W275V | Thermoplasma acidophilum |
| 78.5 | - |
Tm of mutant D176S/S206K/M262I | Thermoplasma acidophilum |
pH Optimum
| pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
|---|---|---|---|
| 7.3 | - |
assay at | Thermoplasma acidophilum |
Cofactor
| Cofactor | Comment | Organism | Structure |
|---|---|---|---|
| additional information | increased to preferred activity of enzyme mutants with NAD+ compared to NADP+ in contrast to wild-type enzyme | Thermoplasma acidophilum | |
| NADP+ | the wild-type enzyme strongly prefers NADP+, negligible activity with NAD+ | Thermoplasma acidophilum | |
Ki Value [mM]
| Ki Value [mM] | Ki Value maximum [mM] | Inhibitor | Comment | Organism | Structure |
|---|---|---|---|---|---|
| 1.25 | - |
D-glyceraldehyde | pH 7.3, temperature not specified in the publication, wild-type enzyme | Thermoplasma acidophilum | |
| 3.8 | - |
D-glyceraldehyde | pH 7.3, temperature not specified in the publication, mutant D176S/S206K/M262I | Thermoplasma acidophilum | |
| 31.2 | - |
D-glyceraldehyde | pH 7.3, temperature not specified in the publication, mutant D176S/S206K/M262I/W271Y/W275V | Thermoplasma acidophilum | |
General Information
| General Information | Comment | Organism |
|---|---|---|
| additional information | structural modeling and docking, the three-dimensional structure of wild-type TaALDH is modeled using the available crystal structure of the triple mutant TaALDH F34M/Y399C/S405N (PDB ID 5M4X) as a template, molecular dynamics simulations | Thermoplasma acidophilum |
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