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Literature summary for 1.5.1.40 extracted from
- Characterization of an 8-hydroxy-5-deazaflavin:NADPH oxidoreductase from Streptomyces griseus (1989), Biochim. Biophys. Acta, 990, 80-86.
General Stability
| General Stability | Organism |
|---|---|
| in 0.08 M NaCl a half-life of 9 h is found which increased to greater than 6000 h in 1 M NaCl (both in the presence of 2% ethylene glycol) | Streptomyces griseus |
KM Value [mM]
| KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|
| 0.0034 | - |
coenzyme F420 | pH 6.0, 22°C | Streptomyces griseus |  |
| 0.0057 | - |
coenzyme F0 | pH 6.0, 22°C | Streptomyces griseus | |
| 0.0129 | - |
reduced coenzyme F420 | pH 6.0, 22°C | Streptomyces griseus | |
| 0.0135 | - |
5'-O-methyl-7,8-didemethyl-8-hydroxyflavin | pH 6.0, 22°C | Streptomyces griseus | |
| 0.0144 | - |
NADP+ | pH 6.0, 22°C | Streptomyces griseus | |
| 0.0155 | - |
7,8-didemethyl-8-hydroxy-5-deazariboflavin 5'-phosphate | pH 6.0, 22°C | Streptomyces griseus | |
| 0.0193 | - |
5-deaza-8-hydroxyisoalloxazine | pH 6.0, 22°C | Streptomyces griseus | |
| 0.0195 | - |
NADPH | pH 6.0, 22°C | Streptomyces griseus | |
| 0.0775 | - |
8-hydroxypyrimido[4,5-b]-2,4-(3H,10H)-dione | pH 6.0, 22°C | Streptomyces griseus | |
Metals/Ions
| Metals/Ions | Comment | Organism | Structure |
|---|---|---|---|
| NaCl | maximal activity at 0.32 M | Streptomyces griseus | |
Molecular Weight [Da]
| Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
|---|---|---|---|
| 28000 | - |
2 * 28000, SDS-PAGE | Streptomyces griseus |
| 42000 | - |
gel filtration | Streptomyces griseus |
Natural Substrates/ Products (Substrates)
| Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| coenzyme F420 + NADPH + H+ | Streptomyces griseus | the main function of this oxidoreductase is probably to provide cells with reduced 8-hydroxy-5-deazaflavin to be used in specific reduction reactions | reduced coenzyme F420 + NADP+ | - |
? | |
Organic Solvent Stability
| Organic Solvent | Comment | Organism |
|---|---|---|
| Ethylene glycol | enhanced the stability: half-life increased from 9 to 750 h in 18% ethylene glycol (in 0.08 M NaCl) | Streptomyces griseus |
| Tween-80 | 0.17% decreases half-life by a factor of approximately 3 | Streptomyces griseus |
Organism
| Organism | UniProt | Comment | Textmining |
|---|---|---|---|
| Streptomyces griseus | - |
- |
- |
Purification (Commentary)
| Purification (Comment) | Organism |
|---|---|
- |
Streptomyces griseus |
Specific Activity [micromol/min/mg]
| Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
|---|---|---|---|
| 11.15 | - |
pH 6.0, 22°C | Streptomyces griseus |
Storage Stability
| Storage Stability | Organism |
|---|---|
| 4°C, activity in crude extract is stable for a month | Streptomyces griseus |
Substrates and Products (Substrate)
| Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| 5'-O-methyl-7,8-didemethyl-8-hydroxyflavin + NADPH + H+ | the enzyme has an absolute requirement for both the 5-deazaflavin structure and the presence of an 8-hydroxy group in the substrate | Streptomyces griseus | 8-hydroxypyrimido[4,5-b]-2,4-(3H,10H)-dione + NADP+ | - |
r | |
| 5-deaza-8-hydroxyisoalloxazine + NADPH + H+ | the enzyme has an absolute requirement for both the 5-deazaflavin structure and the presence of an 8-hydroxy group in the substrate | Streptomyces griseus | 8-hydroxypyrimido[4,5-b]-2,4-(3H,10H)-dione + NADP+ | - |
? | |
| 7,8-didemethyl-8-hydroxy-5-deazariboflavin 5'-phosphate + NADPH + H+ | the enzyme has an absolute requirement for both the 5-deazaflavin structure and the presence of an 8-hydroxy group in the substrate | Streptomyces griseus | 1-deoxy-1-(8-hydroxy-2,4-dioxo-1,3,4,5-tetrahydropyrimido[4,5-b]quinolin-10(2H)-yl)-5-O-phospho-D-ribitol + NADP+ | - |
? | |
| coenzyme F0 + NADPH + H+ | i.e. 7,8-didemethyl-8-hydroxy-5-deazariboflavin. The enzyme has an absolute requirement for both the 5-deazaflavin structure and the presence of an 8-hydroxy group in the substrate | Streptomyces griseus | reduced coenzyme F0 + NADP+ | - |
? | |
| coenzyme F420 + NADPH + H+ | the main function of this oxidoreductase is probably to provide cells with reduced 8-hydroxy-5-deazaflavin to be used in specific reduction reactions | Streptomyces griseus | reduced coenzyme F420 + NADP+ | - |
? | |
| coenzyme F420 + NADPH + H+ | the enzyme has an absolute requirement for both the 5-deazaflavin structure and the presence of an 8-hydroxy group in the substrate. No activity with NAD+ | Streptomyces griseus | reduced coenzyme F420 + NADP+ | - |
r | |
Subunits
| Subunits | Comment | Organism |
|---|---|---|
| dimer | 2 * 28000, SDS-PAGE | Streptomyces griseus |
Synonyms
| Synonyms | Comment | Organism |
|---|---|---|
| 8-OH-5dFl:NADPH oxidoreductase | - |
Streptomyces griseus |
Temperature Optimum [°C]
| Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
|---|---|---|---|
| 22 | - |
assay at | Streptomyces griseus |
Turnover Number [1/s]
| Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|
| 3 | - |
reduced coenzyme F420 | pH 6.0, 22°C | Streptomyces griseus | |
| 4.4 | - |
5-deaza-8-hydroxyisoalloxazine | pH 6.0, 22°C | Streptomyces griseus | |
| 6.5 | - |
5'-O-methyl-7,8-didemethyl-8-hydroxyflavin | pH 6.0, 22°C | Streptomyces griseus | |
| 10.23 | - |
8-hydroxypyrimido[4,5-b]-2,4-(3H,10H)-dione | pH 6.0, 22°C | Streptomyces griseus | |
| 12.15 | - |
coenzyme F0 | pH 6.0, 22°C | Streptomyces griseus | |
| 17.22 | - |
coenzyme F420 | pH 6.0, 22°C | Streptomyces griseus | |
| 35.57 | - |
7,8-didemethyl-8-hydroxy-5-deazariboflavin 5'-phosphate | pH 6.0, 22°C | Streptomyces griseus | |
pH Optimum
| pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
|---|---|---|---|
| 5.9 | - |
reduction of 8-hydroxy-5-azaflavins | Streptomyces griseus |
| 7.9 | - |
oxidation of 8-hydroxy-5-azaflavins | Streptomyces griseus |
Cofactor
| Cofactor | Comment | Organism | Structure |
|---|---|---|---|
| NADP+ | can not be replaced by NAD+ | Streptomyces griseus | |
| NADPH | can not be replaced by NADH | Streptomyces griseus | |
General Information
| General Information | Comment | Organism |
|---|---|---|
| physiological function | the main function of this oxidoreductase is probably to provide cells with reduced 8-hydroxy-5-deazaflavin to be used in specific reduction reactions. The last step of the tetracycline biosynthesis in Streptomyces aureofaciens in which 5a,11a-dehydrochlortetracycline is reduced to chlortetracycline is 8-hydroxy-5-deazaflavin-dependent, and the reducing equivalents are obtained from NADPH | Streptomyces griseus |
kcat/KM [mM/s]
| kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|
| 23 | - |
5-deaza-8-hydroxyisoalloxazine | pH 6.0, 22°C | Streptomyces griseus | |
| 48 | - |
5'-O-methyl-7,8-didemethyl-8-hydroxyflavin | pH 6.0, 22°C | Streptomyces griseus | |
| 132 | - |
8-hydroxypyrimido[4,5-b]-2,4-(3H,10H)-dione | pH 6.0, 22°C | Streptomyces griseus | |
| 233 | - |
reduced coenzyme F420 | pH 6.0, 22°C | Streptomyces griseus | |
| 2132 | - |
coenzyme F0 | pH 6.0, 22°C | Streptomyces griseus | |
| 2295 | - |
7,8-didemethyl-8-hydroxy-5-deazariboflavin 5'-phosphate | pH 6.0, 22°C | Streptomyces griseus | |
| 5065 | - |
coenzyme F420 | pH 6.0, 22°C | Streptomyces griseus | |
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