Any feedback?
Literature summary for 1.1.1.2 extracted from
- Engineering an alcohol dehydrogenase for balancing kinetics in NADPH regeneration with 1,4-butanediol as a cosubstrate (2019), ACS Sust. Chem. Eng., 7, 15706-15714 .
No PubMed abstract available
Application
| Application | Comment | Organism |
|---|---|---|
| synthesis | engineering ADHs for regenerating NADPH by oxidation of diols | Vanderwaltozyma polyspora |
Cloned(Commentary)
| Cloned (Comment) | Organism |
|---|---|
| recombinant expression of His-tagged wild-type and mutant enzymes in Escherichia coli strain BL(DE3) | Vanderwaltozyma polyspora |
Protein Variants
| Protein Variants | Comment | Organism |
|---|---|---|
| F197W | site-directed mutagenesis, kinetics compared to the wild-type enzyme | Vanderwaltozyma polyspora |
| additional information | engineering an alcohol dehydrogenase for balancing kinetics in NADPH regeneration with 1,4-butanediol as a cosubstrate. Identification of the key residues of KpADH by molecular dynamics simulations | Vanderwaltozyma polyspora |
| S196A | site-directed mutagenesis, kinetics compared to the wild-type enzyme | Vanderwaltozyma polyspora |
| S196A/F197W | site-directed mutagenesis, kinetics compared to the wild-type enzyme | Vanderwaltozyma polyspora |
| V84I | site-directed mutagenesis, kinetics compared to the wild-type enzyme | Vanderwaltozyma polyspora |
| V84I/Y127M | site-directed mutagenesis, mutant KpADHV84I/Y127M exhibits a lower KM of 15.1 mM and a higher kcat of 30.1/ min than wild-type KpADH. The ratio of kcat/KM toward 2-hydroxytetrahydrofuran (2-HTHF), in comparison to 1,4-BD, in KpADHV84I/Y127M is dramatically reduced by almost 100fold compared to wild-type KpADH, which is advantageous for NADPH regeneration | Vanderwaltozyma polyspora |
| V84I/Y12C | site-directed mutagenesis, kinetics compared to the wild-type enzyme | Vanderwaltozyma polyspora |
| Y127C | site-directed mutagenesis, kinetics compared to the wild-type enzyme | Vanderwaltozyma polyspora |
| Y127C/S196A | site-directed mutagenesis, kinetics compared to the wild-type enzyme | Vanderwaltozyma polyspora |
| Y127M | site-directed mutagenesis, kinetics compared to the wild-type enzyme | Vanderwaltozyma polyspora |
KM Value [mM]
| KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|
| additional information | - |
additional information | kinetic analysis of recombinant wild-type an d mutant enzymes, overview | Vanderwaltozyma polyspora | |
| 0.0648 | - |
NADP+ | pH 9.5, 30°C, recombinant wild-type enzyme, with 2-hydroxytetrahydrofuran | Vanderwaltozyma polyspora |  |
| 0.0768 | - |
NADP+ | pH 9.5, 30°C, recombinant mutant V127M, with 2-hydroxytetrahydrofuran | Vanderwaltozyma polyspora | |
| 0.0807 | - |
NADP+ | pH 9.5, 30°C, recombinant mutant V84I/Y127M, with 2-hydroxytetrahydrofuran | Vanderwaltozyma polyspora | |
| 0.1 | - |
NADP+ | pH 9.5, 30°C, recombinant mutant S196A/F197W, with 2-hydroxytetrahydrofuran | Vanderwaltozyma polyspora | |
| 0.107 | - |
NADP+ | pH 9.5, 30°C, recombinant mutant F197W, with 2-hydroxytetrahydrofuran | Vanderwaltozyma polyspora | |
| 0.107 | - |
NADP+ | pH 9.5, 30°C, recombinant mutant V127C, with 2-hydroxytetrahydrofuran | Vanderwaltozyma polyspora | |
| 0.159 | - |
NADP+ | pH 9.5, 30°C, recombinant mutant V84I, with 2-hydroxytetrahydrofuran | Vanderwaltozyma polyspora | |
| 0.456 | - |
NADP+ | pH 9.5, 30°C, recombinant mutant S196A, with 2-hydroxytetrahydrofuran | Vanderwaltozyma polyspora | |
| 0.462 | - |
NADP+ | pH 9.5, 30°C, recombinant mutant V84I/Y127C, with 2-hydroxytetrahydrofuran | Vanderwaltozyma polyspora | |
| 0.565 | - |
NADP+ | pH 9.5, 30°C, recombinant mutant Y127C/S196A, with 2-hydroxytetrahydrofuran | Vanderwaltozyma polyspora | |
| 0.85 | - |
2-hydroxytetrahydrofuran | pH 9.5, 30°C, recombinant mutant S196A/F197W | Vanderwaltozyma polyspora | |
| 1.55 | - |
2-hydroxytetrahydrofuran | pH 9.5, 30°C, recombinant mutant S196A | Vanderwaltozyma polyspora | |
| 1.59 | - |
2-hydroxytetrahydrofuran | pH 9.5, 30°C, recombinant mutant F197W | Vanderwaltozyma polyspora | |
| 1.9 | - |
2-hydroxytetrahydrofuran | pH 9.5, 30°C, recombinant mutant V84I | Vanderwaltozyma polyspora | |
| 2.23 | - |
2-hydroxytetrahydrofuran | pH 9.5, 30°C, recombinant wild-type enzyme | Vanderwaltozyma polyspora | |
| 2.4 | - |
2-hydroxytetrahydrofuran | pH 9.5, 30°C, recombinant mutant V127C | Vanderwaltozyma polyspora | |
| 4.39 | - |
2-hydroxytetrahydrofuran | pH 9.5, 30°C, recombinant mutant V84I/Y127M | Vanderwaltozyma polyspora | |
| 4.7 | - |
2-hydroxytetrahydrofuran | pH 9.5, 30°C, recombinant mutant V84I/Y127C | Vanderwaltozyma polyspora | |
| 5.9 | - |
2-hydroxytetrahydrofuran | pH 9.5, 30°C, recombinant mutant V127M | Vanderwaltozyma polyspora | |
| 6.38 | - |
2-hydroxytetrahydrofuran | pH 9.5, 30°C, recombinant mutant Y127C/S196A | Vanderwaltozyma polyspora | |
| 15.1 | - |
1,4-Butanediol | pH 9.5, 30°C, recombinant mutant V84I/Y127M | Vanderwaltozyma polyspora | |
Organism
| Organism | UniProt | Comment | Textmining |
|---|---|---|---|
| Vanderwaltozyma polyspora | - |
- |
- |
Purification (Commentary)
| Purification (Comment) | Organism |
|---|---|
| recombinant His-tagged wild-type and mutant enzymes from Escherichia coli strain BL(DE3) by nickel affinity chromatography | Vanderwaltozyma polyspora |
Substrates and Products (Substrate)
| Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| 1,4-butanediol + NADP+ | - |
Vanderwaltozyma polyspora | 4-hydroxybutanal + NADPH + H+ | - |
r | |
| 1,5-pentanediol + NADP+ | - |
Vanderwaltozyma polyspora | 5-hydroxypentanal + NADPH + H+ | - |
r | |
| 1,6-hexanediol + NADP+ | - |
Vanderwaltozyma polyspora | 6-hydroxyhexanal + NADPH + H+ | - |
r | |
| 2-hydroxytetrahydrofuran + NADP+ | - |
Vanderwaltozyma polyspora | 2-oxotetrahydrofuran + NADPH + H+ | - |
ir | |
| additional information | the oxidation of 1,4-BD to 4-hydroxybutanal is the first, reversible, rate-limiting step in the NADPH regeneration system with 1,4-butanediol (1,4-BD). It is followed by spontaneous formation of 2-hydroxytetrahydrofuran (2-HTHF) from 4-hydroxybutanal via cyclization. The second enzymatic step, the reduction of 2-HTHF into gamma-butyrolactone, is irreversible, liberating 2 equiv. of NAD(P)H | Vanderwaltozyma polyspora | ? | - |
- |
|
Synonyms
| Synonyms | Comment | Organism |
|---|---|---|
| KpADH | - |
Vanderwaltozyma polyspora |
| NADP+-dependent alcohol dehydrogenase | - |
Vanderwaltozyma polyspora |
Temperature Optimum [°C]
| Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
|---|---|---|---|
| 30 | - |
- |
Vanderwaltozyma polyspora |
Turnover Number [1/s]
| Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|
| 0.000038 | - |
2-hydroxytetrahydrofuran | pH 9.5, 30°C, recombinant mutant S196A | Vanderwaltozyma polyspora | |
| 0.000053 | - |
2-hydroxytetrahydrofuran | pH 9.5, 30°C, recombinant mutant V84I/Y127C | Vanderwaltozyma polyspora | |
| 0.000062 | - |
2-hydroxytetrahydrofuran | pH 9.5, 30°C, recombinant mutant Y127C/S196A | Vanderwaltozyma polyspora | |
| 0.000074 | - |
2-hydroxytetrahydrofuran | pH 9.5, 30°C, recombinant mutant S196A/F197W | Vanderwaltozyma polyspora | |
| 0.000077 | - |
2-hydroxytetrahydrofuran | pH 9.5, 30°C, recombinant mutant V127C | Vanderwaltozyma polyspora | |
| 0.00014 | - |
2-hydroxytetrahydrofuran | pH 9.5, 30°C, recombinant mutant V84I/Y127M | Vanderwaltozyma polyspora | |
| 0.00018 | - |
2-hydroxytetrahydrofuran | pH 9.5, 30°C, recombinant mutant F197W | Vanderwaltozyma polyspora | |
| 0.00022 | - |
2-hydroxytetrahydrofuran | pH 9.5, 30°C, recombinant mutant V84I | Vanderwaltozyma polyspora | |
| 0.00039 | - |
2-hydroxytetrahydrofuran | pH 9.5, 30°C, recombinant mutant V127M | Vanderwaltozyma polyspora | |
| 0.00044 | - |
2-hydroxytetrahydrofuran | pH 9.5, 30°C, recombinant wild-type enzyme | Vanderwaltozyma polyspora | |
| 0.502 | - |
1,4-Butanediol | pH 9.5, 30°C, mutant V84I/Y127M | Vanderwaltozyma polyspora | |
pH Optimum
| pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
|---|---|---|---|
| 9.5 | - |
- |
Vanderwaltozyma polyspora |
Cofactor
| Cofactor | Comment | Organism | Structure |
|---|---|---|---|
| NADP+ | - |
Vanderwaltozyma polyspora | |
| NADPH | - |
Vanderwaltozyma polyspora | |
kcat/KM [mM/s]
| kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|
| 0.0000032 | - |
2-hydroxytetrahydrofuran | pH 9.5, 30°C, recombinant mutant V127C | Vanderwaltozyma polyspora | |
| 0.0000066 | - |
2-hydroxytetrahydrofuran | pH 9.5, 30°C, recombinant mutant V127M | Vanderwaltozyma polyspora | |
| 0.0000097 | - |
2-hydroxytetrahydrofuran | pH 9.5, 30°C, recombinant mutant Y127C/S196A | Vanderwaltozyma polyspora | |
| 0.000011 | - |
2-hydroxytetrahydrofuran | pH 9.5, 30°C, recombinant mutant V84I/Y127C | Vanderwaltozyma polyspora | |
| 0.000012 | - |
2-hydroxytetrahydrofuran | pH 9.5, 30°C, recombinant mutant V84I | Vanderwaltozyma polyspora | |
| 0.000032 | - |
2-hydroxytetrahydrofuran | pH 9.5, 30°C, recombinant mutant V84I/Y127M | Vanderwaltozyma polyspora | |
| 0.000087 | - |
2-hydroxytetrahydrofuran | pH 9.5, 30°C, recombinant mutant S196A/F197W | Vanderwaltozyma polyspora | |
| 0.00011 | - |
2-hydroxytetrahydrofuran | pH 9.5, 30°C, recombinant mutant F197W | Vanderwaltozyma polyspora | |
| 0.0002 | - |
2-hydroxytetrahydrofuran | pH 9.5, 30°C, recombinant wild-type enzyme | Vanderwaltozyma polyspora | |
| 0.00025 | - |
2-hydroxytetrahydrofuran | pH 9.5, 30°C, recombinant mutant S196A | Vanderwaltozyma polyspora | |
| 0.033 | - |
1,4-Butanediol | pH 9.5, 30°C, mutant V84I/Y127M | Vanderwaltozyma polyspora | |
Use of this online version of BRENDA is free under the CC BY 4.0 license. See terms of use for full details.
Release 2023.1 (January 2023)
Legal
Curated at
Member of
