Cloned (Comment) | Organism |
---|---|
gene bld, butyraldehyde dehydrogenase gene of Clostridium saccharoperbutylacetonicum is coexpresses with diverse involved genes in Escherichia coli for the reconstruction of 1,4-butanediol biosynthesis pathway in Escherichia coli, recombinant expression of His-tagged wild-type and mutant enzymes in Escherichia coli strain BL21(DE3) | Clostridium saccharoperbutylacetonicum |
Protein Variants | Comment | Organism |
---|---|---|
A176T | random mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme | Clostridium saccharoperbutylacetonicum |
A467S | random mutagenesis, the mutant shows similar activity compared to the wild-type enzyme | Clostridium saccharoperbutylacetonicum |
K279R | random mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme | Clostridium saccharoperbutylacetonicum |
L273C | site-directed mutagenesis, the mutant shows about 2fold increased activity compared to the wild-type enzyme | Clostridium saccharoperbutylacetonicum |
L273I | random mutagenesis, the mutant shows about 3.5fold increased activity compared to the wild-type enzyme | Clostridium saccharoperbutylacetonicum |
L273M | site-directed mutagenesis, the mutant shows about 2fold increased activity compared to the wild-type enzyme | Clostridium saccharoperbutylacetonicum |
L273S | site-directed mutagenesis, the mutant shows slightly increased activity compared to the wild-type enzyme | Clostridium saccharoperbutylacetonicum |
L273T | site-directed mutagenesis, the mutant shows about 4fold increased activity compared to the wild-type enzyme | Clostridium saccharoperbutylacetonicum |
L273V | site-directed mutagenesis, the mutant shows slightly increased activity compared to the wild-type enzyme | Clostridium saccharoperbutylacetonicum |
M371R | random mutagenesis, the mutant shows highly reduced activity compared to the wild-type enzyme | Clostridium saccharoperbutylacetonicum |
additional information | to fit enzyme Bld into the non-natural 1,4-butanediol pathway, it is engineered through random mutagenesis, library construction. Five Bld mutants are isolated using a colorimetric Schiff's reagent-based method. Subsequent site-directed mutagenesis of Bld generates the two best Bld mutants, L273I and L273T, which produce 1,4-butanediol titers fourfold greater than those of wild-type Bld. Butyraldehyde dehydrogenase (Bld) and butanol dehydrogenase (Bdh) of Clostridium saccharoperbutylacetonicum are selected as a substitute for the bifunctional AdhE2 in the reconstructed 1,4-butanediol biosynthesis pathway in Escherichia coli, coexpression of diverse involved genes in Escherichia coli | Clostridium saccharoperbutylacetonicum |
N409T | random mutagenesis, the mutant shows similar activity compared to the wild-type enzyme | Clostridium saccharoperbutylacetonicum |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
additional information | - |
additional information | no Michaelis-Menten behaviour | Clostridium saccharoperbutylacetonicum |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
4-hydroxybutanoyl-CoA + NADH + H+ | Clostridium saccharoperbutylacetonicum | - |
4-hydroxybutanal + CoA + NAD+ | - |
? | |
4-hydroxybutanoyl-CoA + NADH + H+ | Clostridium saccharoperbutylacetonicum KCTC5577 | - |
4-hydroxybutanal + CoA + NAD+ | - |
? | |
4-hydroxybutanoyl-CoA + NADPH + H+ | Clostridium saccharoperbutylacetonicum | - |
4-hydroxybutanal + CoA + NADP+ | - |
? | |
4-hydroxybutanoyl-CoA + NADPH + H+ | Clostridium saccharoperbutylacetonicum KCTC5577 | - |
4-hydroxybutanal + CoA + NADP+ | - |
? | |
butanoyl-CoA + NADH + H+ | Clostridium saccharoperbutylacetonicum | - |
butanal + CoA + NAD+ | - |
? | |
butanoyl-CoA + NADH + H+ | Clostridium saccharoperbutylacetonicum KCTC5577 | - |
butanal + CoA + NAD+ | - |
? | |
butanoyl-CoA + NADPH + H+ | Clostridium saccharoperbutylacetonicum | - |
butanal + CoA + NADP+ | - |
? | |
butanoyl-CoA + NADPH + H+ | Clostridium saccharoperbutylacetonicum KCTC5577 | - |
butanal + CoA + NADP+ | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Clostridium saccharoperbutylacetonicum | - |
- |
- |
Clostridium saccharoperbutylacetonicum KCTC5577 | - |
- |
- |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
4-hydroxybutanoyl-CoA + NADH + H+ | - |
Clostridium saccharoperbutylacetonicum | 4-hydroxybutanal + CoA + NAD+ | - |
? | |
4-hydroxybutanoyl-CoA + NADH + H+ | - |
Clostridium saccharoperbutylacetonicum KCTC5577 | 4-hydroxybutanal + CoA + NAD+ | - |
? | |
4-hydroxybutanoyl-CoA + NADPH + H+ | - |
Clostridium saccharoperbutylacetonicum | 4-hydroxybutanal + CoA + NADP+ | - |
? | |
4-hydroxybutanoyl-CoA + NADPH + H+ | - |
Clostridium saccharoperbutylacetonicum KCTC5577 | 4-hydroxybutanal + CoA + NADP+ | - |
? | |
butanoyl-CoA + NADH + H+ | - |
Clostridium saccharoperbutylacetonicum | butanal + CoA + NAD+ | - |
? | |
butanoyl-CoA + NADH + H+ | - |
Clostridium saccharoperbutylacetonicum KCTC5577 | butanal + CoA + NAD+ | - |
? | |
butanoyl-CoA + NADPH + H+ | - |
Clostridium saccharoperbutylacetonicum | butanal + CoA + NADP+ | - |
? | |
butanoyl-CoA + NADPH + H+ | - |
Clostridium saccharoperbutylacetonicum KCTC5577 | butanal + CoA + NADP+ | - |
? |
Synonyms | Comment | Organism |
---|---|---|
Bld | - |
Clostridium saccharoperbutylacetonicum |
Butyraldehyde dehydrogenase | - |
Clostridium saccharoperbutylacetonicum |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
NADH | - |
Clostridium saccharoperbutylacetonicum | |
NADPH | - |
Clostridium saccharoperbutylacetonicum |
General Information | Comment | Organism |
---|---|---|
metabolism | the enzyme is part of the 1,4-butanediol biosynthesis pathway, overview | Clostridium saccharoperbutylacetonicum |