Cloned (Comment) | Organism |
---|---|
gene bldh, recombinant expression in Escherichia coli strain JCL166, strain JCL166 cannot grow anaerobically unless complemented by an exogenous fermentation pathway such as n-butanol biosynthesis | Synechococcus elongatus |
gene bldh, recombinant expression in Escherichia coli strain JCL166, strain JCL166 cannot grow anaerobically unless complemented by an exogenous fermentation pathway such as n-butanol biosynthesis. Recombinant coexpression of PduP with the enzymes of the n-butanol synthesis pathway in Synechococcus elongatus strain PCC 7942 results in autotrophic n-butanol production | Clostridium saccharoperbutylacetonicum |
gene pduP, recombinant expression in Escherichia coli strain JCL166, strain JCL166 cannot grow anaerobically unless complemented by an exogenous fermentation pathway such as n-butanol biosynthesis. Recombinant coexpression of PduP with the enzymes of the n-butanol synthesis pathway in Synechococcus elongatus strain PCC 7942 results in autotrophic n-butanol production. PduP from Lactobacillus brevis produces more n-butanol than ethanol | Levilactobacillus brevis |
Protein Variants | Comment | Organism |
---|---|---|
additional information | design of a coenzyme A (CoA) dependent n-butanol biosynthesis pathway tailored to the metabolic physiology of the cyanobacterium Synechococcus elongatus PCC 7942 by incorporating an ATP driving force and a kinetically irreversible trap. Oxygen-sensitive CoA-acylating butyraldehyde dehydrogenase (Bldh) is exchanged for the oxygen-tolerant PduP from Salmonella enterica. Replacing Bldh with PduP in the n-butanol synthesis pathway results in n-butanol production to a cumulative titer of 404 mg/l with peak productivity of 51 mg/l per day, exceeding the base strain by 20fold. Anaerobic growth rescue of Escherichia coli strain JCL166 by overexpression of the Clostridium butanol pathway with different aldehyde dehydrogenases PduP | Synechococcus elongatus |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
additional information | - |
additional information | no Michaelis-Menten behaviour | Clostridium saccharoperbutylacetonicum | |
0.076 | - |
acetyl-CoA | recombinant enzyme, pH 7.15, 30°C | Levilactobacillus brevis | |
0.534 | - |
butanoyl-CoA | recombinant enzyme, pH 7.15, 30°C | Levilactobacillus brevis |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
acetyl-CoA + NADH + H+ | Levilactobacillus brevis | - |
acetaldehyde + CoA + NAD+ | - |
r | |
acetyl-CoA + NADH + H+ | Clostridium saccharoperbutylacetonicum | low activity | acetaldehyde + CoA + NAD+ | - |
r | |
butanoyl-CoA + NADH + H+ | Levilactobacillus brevis | - |
butanal + CoA + NAD+ | - |
r | |
butanoyl-CoA + NADH + H+ | Synechococcus elongatus | - |
butanal + CoA + NAD+ | - |
? | |
butanoyl-CoA + NADH + H+ | Clostridium saccharoperbutylacetonicum | best substrate | butanal + CoA + NAD+ | - |
r | |
butanoyl-CoA + NADH + H+ | Synechococcus elongatus PCC 7942 | - |
butanal + CoA + NAD+ | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Clostridium saccharoperbutylacetonicum | - |
- |
- |
Levilactobacillus brevis | - |
- |
- |
Synechococcus elongatus | - |
- |
- |
Synechococcus elongatus PCC 7942 | - |
- |
- |
Oxidation Stability | Organism |
---|---|
oxygen sensitivity of CoA-acylating aldehyde dehydrogenase | Synechococcus elongatus |
the enzyme is oxygen-tolerant | Levilactobacillus brevis |
the enzyme is oxygen-tolerant | Clostridium saccharoperbutylacetonicum |
Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
---|---|---|---|
0.49 | - |
with butanoyl-CoA, pH 7.15, 30°C | Clostridium saccharoperbutylacetonicum |
2.5 | - |
with butanoyl-CoA, pH 7.15, 30°C | Levilactobacillus brevis |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
acetyl-CoA + NADH + H+ | - |
Levilactobacillus brevis | acetaldehyde + CoA + NAD+ | - |
r | |
acetyl-CoA + NADH + H+ | low activity | Clostridium saccharoperbutylacetonicum | acetaldehyde + CoA + NAD+ | - |
r | |
butanoyl-CoA + NADH + H+ | - |
Levilactobacillus brevis | butanal + CoA + NAD+ | - |
r | |
butanoyl-CoA + NADH + H+ | - |
Synechococcus elongatus | butanal + CoA + NAD+ | - |
? | |
butanoyl-CoA + NADH + H+ | best substrate | Clostridium saccharoperbutylacetonicum | butanal + CoA + NAD+ | - |
r | |
butanoyl-CoA + NADH + H+ | - |
Synechococcus elongatus PCC 7942 | butanal + CoA + NAD+ | - |
? | |
additional information | substrate chain length specificity of the enzyme is C2-C12, highest activity with C6 substrate, overview | Levilactobacillus brevis | ? | - |
? |
Synonyms | Comment | Organism |
---|---|---|
Bldh | - |
Synechococcus elongatus |
Bldh | - |
Clostridium saccharoperbutylacetonicum |
CoA-acylating aldehyde dehydrogenase | - |
Levilactobacillus brevis |
CoA-acylating aldehyde dehydrogenase | - |
Synechococcus elongatus |
CoA-acylating aldehyde dehydrogenase | - |
Clostridium saccharoperbutylacetonicum |
CoA-acylating butyraldehyde dehydrogenase | - |
Synechococcus elongatus |
coenzyme A-acylating aldehyde dehydrogenase | - |
Levilactobacillus brevis |
coenzyme A-acylating aldehyde dehydrogenase | - |
Synechococcus elongatus |
coenzyme A-acylating aldehyde dehydrogenase | - |
Clostridium saccharoperbutylacetonicum |
PduP | - |
Levilactobacillus brevis |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
30 | - |
assay at | Levilactobacillus brevis |
30 | - |
assay at | Clostridium saccharoperbutylacetonicum |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.26 | - |
acetyl-CoA | recombinant enzyme, pH 7.15, 30°C | Levilactobacillus brevis | |
3.37 | - |
butanoyl-CoA | recombinant enzyme, pH 7.15, 30°C | Levilactobacillus brevis |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
7.15 | - |
assay at | Levilactobacillus brevis |
7.15 | - |
assay at | Clostridium saccharoperbutylacetonicum |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
NADH | - |
Levilactobacillus brevis | |
NADH | - |
Synechococcus elongatus | |
NADH | - |
Clostridium saccharoperbutylacetonicum |
General Information | Comment | Organism |
---|---|---|
metabolism | the oxygen sensitivity of CoA-acylating aldehyde dehydrogenase appears to be a key limiting factor for cyanobacteria to produce alcohols through the CoA-dependent route | Levilactobacillus brevis |
metabolism | the oxygen sensitivity of CoA-acylating aldehyde dehydrogenase appears to be a key limiting factor for cyanobacteria to produce alcohols through the CoA-dependent route | Synechococcus elongatus |
metabolism | the oxygen sensitivity of CoA-acylating aldehyde dehydrogenase appears to be a key limiting factor for cyanobacteria to produce alcohols through the CoA-dependent route | Clostridium saccharoperbutylacetonicum |
kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
3 | - |
acetyl-CoA | recombinant enzyme, pH 7.15, 30°C | Levilactobacillus brevis | |
6 | - |
butanoyl-CoA | recombinant enzyme, pH 7.15, 30°C | Levilactobacillus brevis |