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Literature summary extracted from
- Cloning and expression of a Clostridium kluyveri gene responsible for diaphorase activity (2008), Biosci. Biotechnol. Biochem., 72, 735-741.
Cloned(Commentary)
| EC Number | Cloned (Comment) | Organism |
|---|---|---|
| 1.8.1.4 | expressed in Escherichia coli as a recombinant protein | Clostridium kluyveri |
KM Value [mM]
| EC Number | KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|---|
| 1.8.1.4 | 0.09 | - |
NADH | native DiaA | Clostridium kluyveri |  |
| 1.8.1.4 | 0.15 | - |
NADH | recombinant DiaA | Clostridium kluyveri | |
| 1.8.1.4 | 0.35 | - |
NADPH | recombinant DiaA | Clostridium kluyveri | |
| 1.8.1.4 | 0.5 | - |
nitrotetrazolium blue | recombinant DiaA | Clostridium kluyveri | |
Metals/Ions
| EC Number | Metals/Ions | Comment | Organism | Structure |
|---|---|---|---|---|
| 1.8.1.4 | Fe | amino acid sequence homology indicates the presence of a Fe-containing rubredoxin-like domain. The amount of iron is determined to be 1.1 molecule per rDiaA molecule by ICP atomic emission spectroscopy | Clostridium kluyveri | |
Molecular Weight [Da]
| EC Number | Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
|---|---|---|---|---|
| 1.8.1.4 | 18000 | - |
native protein using SDS-PAGE | Clostridium kluyveri |
| 1.8.1.4 | 24980 | - |
predicted from cDNA | Clostridium kluyveri |
| 1.8.1.4 | 28000 | - |
recombinant protein using SDS-PAGE, in good agreement with the calculated molecular weight of recombinant DiaA | Clostridium kluyveri |
| 1.8.1.4 | 86000 | - |
native form of recombinant DiaA, gel filtration | Clostridium kluyveri |
Organism
| EC Number | Organism | UniProt | Comment | Textmining |
|---|---|---|---|---|
| 1.8.1.4 | Clostridium kluyveri | - |
- |
- |
Purification (Commentary)
| EC Number | Purification (Comment) | Organism |
|---|---|---|
| 1.8.1.4 | enzmye from culture supernatant of Clostridium kluyveri is purified using a DEAE-toyopearl column and a TSK-GEL G-3000 column. Recombinant protein is purified using a HiTrap chelating HP column | Clostridium kluyveri |
Source Tissue
| EC Number | Source Tissue | Comment | Organism | Textmining |
|---|
Substrates and Products (Substrate)
| EC Number | Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
|---|---|---|---|---|---|---|---|
| 1.8.1.4 | NADPH + H+ + oxidized 2,6-dichlorophenolindophenol | - |
Clostridium kluyveri | NADP+ + reduced 2,6-dichlorophenolindophenol | - |
? | |
| 1.8.1.4 | nitrotetrazolium blue + NADH + H+ | - |
Clostridium kluyveri | reduced nitrotetrazolium blue + NAD+ | - |
r | |
| 1.8.1.4 | oxidized 2,6-dichlorophenolindophenol + NADH | - |
Clostridium kluyveri | ? + NAD+ | - |
? | |
Subunits
| EC Number | Subunits | Comment | Organism |
|---|---|---|---|
| 1.8.1.4 | trimer | gel filtration, 3 * 28000 Da | Clostridium kluyveri |
Synonyms
| EC Number | Synonyms | Comment | Organism |
|---|---|---|---|
| 1.8.1.4 | diaphorase | - |
Clostridium kluyveri |
Temperature Optimum [°C]
| EC Number | Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
|---|---|---|---|---|
| 1.8.1.4 | 40 | - |
- |
Clostridium kluyveri |
Temperature Stability [°C]
| EC Number | Temperature Stability Minimum [°C] | Temperature Stability Maximum [°C] | Comment | Organism |
|---|---|---|---|---|
| 1.8.1.4 | 40 | - |
recombinant enzyme is stable at 30°C upon 30 min of incubation without substrates, but incubation at 40°C and higher temperatures inactivated enzyme activity | Clostridium kluyveri |
pH Optimum
| EC Number | pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
|---|---|---|---|---|
| 1.8.1.4 | 8 | - |
- |
Clostridium kluyveri |
pH Range
| EC Number | pH Minimum | pH Maximum | Comment | Organism |
|---|---|---|---|---|
| 1.8.1.4 | 6 | 9 | - |
Clostridium kluyveri |
pH Stability
| EC Number | pH Stability | pH Stability Maximum | Comment | Organism |
|---|---|---|---|---|
| 1.8.1.4 | 6 | 11 | enzyme loses its activity completely below pH 6 and above pH 11 | Clostridium kluyveri |
Cofactor
| EC Number | Cofactor | Comment | Organism | Structure |
|---|---|---|---|---|
| 1.8.1.4 | FMN | - |
Clostridium kluyveri | |
| 1.8.1.4 | NADH | - |
Clostridium kluyveri | |
| 1.8.1.4 | NADPH | - |
Clostridium kluyveri | |
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