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Literature summary for 1.8.1.4 extracted from
- Characterization of a dihydrolipoyl dehydrogenase having diaphorase activity of Clostridium kluyveri (2008), Biosci. Biotechnol. Biochem., 72, 982-988.
Application
| Application | Comment | Organism |
|---|---|---|
| additional information | high stability of rBfmBC may make it useful for practical use | Clostridium kluyveri |
Cloned(Commentary)
| Cloned (Comment) | Organism |
|---|---|
| bfmBC gene encoding DLD. Ligated into vector pCR2.1, and introduced into Escherichia coli DH5alpha. Insert DNA recovered from the recombinant plasmid ligated into vector pET-28a(+), yielding pET-bfmBC and expressed in Escherichia coli BL21 (DE3) | Clostridium kluyveri |
Inhibitors
| Inhibitors | Comment | Organism | Structure |
|---|---|---|---|
| Hg2+ | 1 mM shows strong inhibitory effect on recombinant rBfmBC activity (more than 80% inhibition) | Clostridium kluyveri |  |
| Pb2+ | 1 mM shows strong inhibitory effect on recombinant BfmBC activity (more than 80% inhibition) | Clostridium kluyveri | |
| Zn2+ | about 30% inhibition of recombinant BfmBC activity with 5 mM | Clostridium kluyveri | |
KM Value [mM]
| KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|
| 0.038 | - |
NADH | - |
Clostridium kluyveri | |
| 0.12 | - |
2,6-dichlorophenolindophenol | in diaphorase activity | Clostridium kluyveri | |
| 0.25 | - |
NADH | in diaphorase activity | Clostridium kluyveri | |
| 0.3 | - |
dihydrolipoamide | - |
Clostridium kluyveri | |
| 0.42 | - |
Lipoamide | - |
Clostridium kluyveri | |
| 0.5 | - |
NAD+ | - |
Clostridium kluyveri | |
Metals/Ions
| Metals/Ions | Comment | Organism | Structure |
|---|---|---|---|
| additional information | 1 mM Na+, Ca2+, Mg2+, Mn2+, Fe3+, Zn2+, Pb2+, and Hg2+ do not enhance enzyme activity | Clostridium kluyveri |
Molecular Weight [Da]
| Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
|---|---|---|---|
| 50000 | - |
2 * 50000, SDS-PAGE, 2 * 53000, sequence analysis | Clostridium kluyveri |
| 53000 | - |
2 * 50000, SDS-PAGE, 2 * 53000, sequence analysis | Clostridium kluyveri |
| 110000 | - |
gel filtration | Clostridium kluyveri |
Organism
| Organism | UniProt | Comment | Textmining |
|---|---|---|---|
| Clostridium kluyveri | A5N930 | strain ATCC 8527 | - |
Purification (Commentary)
| Purification (Comment) | Organism |
|---|---|
| by gel filtration | Clostridium kluyveri |
Storage Stability
| Storage Stability | Organism |
|---|---|
| 4°C, 3 months | Clostridium kluyveri |
Substrates and Products (Substrate)
| Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| 2,6-dichlorophenolindophenol + NADH + H+ | - |
Clostridium kluyveri | reduced 2,6-dichlorophenolindophenol + NAD+ | - |
r | |
| dihydrolipoamide + NAD+ | - |
Clostridium kluyveri | lipoamide + NADH | - |
r | |
| lipoamide + NADH | - |
Clostridium kluyveri | dihydrolipoamide + NAD+ | - |
r | |
| nitro blue tetrazolium + NADH | - |
Clostridium kluyveri | ? + NAD+ | - |
? | |
| oxidized 2,6-dichlorophenolindophenol + NADH | - |
Clostridium kluyveri | ? + NAD+ | - |
? | |
Subunits
| Subunits | Comment | Organism |
|---|---|---|
| dimer | 2 * 50000, SDS-PAGE, 2 * 53000, sequence analysis | Clostridium kluyveri |
Synonyms
| Synonyms | Comment | Organism |
|---|---|---|
| DLD | - |
Clostridium kluyveri |
Temperature Optimum [°C]
| Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
|---|---|---|---|
| 40 | - |
- |
Clostridium kluyveri |
Temperature Stability [°C]
| Temperature Stability Minimum [°C] | Temperature Stability Maximum [°C] | Comment | Organism |
|---|---|---|---|
| 60 | - |
the enzyme maintains some activity after a 30-min incubation | Clostridium kluyveri |
pH Optimum
| pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
|---|---|---|---|
| 7 | - |
- |
Clostridium kluyveri |
pH Range
| pH Minimum | pH Maximum | Comment | Organism |
|---|---|---|---|
| 6 | 8 | - |
Clostridium kluyveri |
pH Stability
| pH Stability | pH Stability Maximum | Comment | Organism |
|---|---|---|---|
| 6 | 9 | retains about half of the initial activity | Clostridium kluyveri |
Cofactor
| Cofactor | Comment | Organism | Structure |
|---|---|---|---|
| FAD | contains a noncovalently but tightly attached FAD molecule | Clostridium kluyveri | |
| additional information | FMN is not present in the recombinant enzyme as a cofactor. Does not use NADPH as the electron donor | Clostridium kluyveri | |
| NAD+ | - |
Clostridium kluyveri | |
| NADH | has a preference for NADH as a coenzyme | Clostridium kluyveri | |
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