Cloned (Comment) | Organism |
---|---|
expression in Escherichia coli | Plagiochasma appendiculatum |
Protein Variants | Comment | Organism |
---|---|---|
A120M | when the purified recombinant mutant enzyme is incubated with naringenin as substrate, the ratio of 2-hydroxynaringenin to apigenin is reduced. The mutant enzyme is able to oxidize dihydrokaempferol to kaempferol with 162% of the catalytic activity relative to wild-type enzyme | Plagiochasma appendiculatum |
F146I | when the purified recombinant mutant enzyme is incubated with naringenin as substrate, the ratio of 2-hydroxynaringenin to apigenin is reduced. The mutant enzyme is able to oxidize dihydrokaempferol to kaempferol with 86% of the catalytic activity relative to wild-type enzyme | Plagiochasma appendiculatum |
F146I/A120M | the enzyme activity of the double mutant is reduced to a level of about 10% of the activity of the wild type enzyme and the only product generated is apigenin | Plagiochasma appendiculatum |
L311F | when the purified recombinant mutant enzyme is incubated with naringenin as substrate, the ratio of 2-hydroxynaringenin to apigenin is reduced. The mutant enzyme is able to oxidize dihydrokaempferol to kaempferol with 355% of the catalytic activity relative to wild-type enzyme | Plagiochasma appendiculatum |
L311F/A120M | the double mutant catalyzes the formation of 2-hydroxynaringenin and apigenin, although the ratio of these two compounds iss lower than that generated by the wild type enzyme | Plagiochasma appendiculatum |
L311F/F146I | the double mutant catalyzes the formation of 2-hydroxynaringenin and apigenin, although the ratio of these two compounds iss lower than that generated by the wild type enzyme | Plagiochasma appendiculatum |
L311F/F146I/A120M | the activity of the triple mutant falls to about 5% of the wild type enzyme | Plagiochasma appendiculatum |
L311F/F146I/Y240P | inactive mutant | Plagiochasma appendiculatum |
L311F/F146I/Y240P/A120M | inactive mutant | Plagiochasma appendiculatum |
L311F/Y240P | the double mutant exhibits less flavone synthase activity than either the wild type or the Y240P single mutant enzymes | Plagiochasma appendiculatum |
Y240P | the mutant enzyme converts naringenin to apigenin without producing any 2-hydroxynaringenin. The mutant has a higher affinity (lower Km) with naringenin than the wild type enzyme, but the catalytic efficiency of the mutant is lower than that of wild type enzyme. The mutant enzyme is able to oxidize dihydrokaempferol to kaempferol with 104% of the catalytic activity relative to wild-type enzyme | Plagiochasma appendiculatum |
Y240P/A120M | the enzyme activity of the double mutant is reduced to a level of about 10% of the activity of the wild type enzyme and the only product generated is apigenin. The mutant enzyme is able to oxidize dihydrokaempferol to kaempferol with 175% of the catalytic activity relative to wild-type enzyme | Plagiochasma appendiculatum |
Y240P/F146I | the enzyme activity of the double mutant is reduced to a level of about 10% of the activity of the wild type enzyme and the only product generated is apigenin | Plagiochasma appendiculatum |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.0065 | - |
naringenin | pH 7.0, 35°C, mutant enzyme Y240P | Plagiochasma appendiculatum | |
0.0199 | - |
naringenin | pH 7.0, 35°C, wild-type enzyme | Plagiochasma appendiculatum |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Plagiochasma appendiculatum | A0A076U8J8 | - |
- |
Purification (Comment) | Organism |
---|---|
- |
Plagiochasma appendiculatum |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
dihydrokaempferol + 2-oxoglutarate + O2 | the mutated enzymes L311F, Y240P, F146I, A120M and L311FA120M are all able to oxidize dihydrokaempferol to kaempferol, with respective catalytic activities relative to the wild type level of 355%, 104%, 86%, 162% and 175%. L311F substitution increases the flavonol synthase activity | Plagiochasma appendiculatum | kaempferol + succinate + CO2 + H2O | - |
? | |
additional information | no activity with eriodictyol or dihydroquercetin | Plagiochasma appendiculatum | ? | - |
? | |
naringenin + 2-oxoglutarate + O2 | the enzyme catalyzes the conversion of naringenin to apigenin and 2-hydroxynaringenin. It is demonstrated that the enzyme is unable to catalyze the conversion of 2-hydroxynaringenin to apigenin. It is also confirmed that 2-hydroxynaringenin cannot be an intermediate product in the conversion of naringenin to apigenin, leaving the mechanism of conversion unknown | Plagiochasma appendiculatum | apigenin + 2-hydroxynaringenin + succinate + CO2 + H2O | - |
? |
Subunits | Comment | Organism |
---|---|---|
? | x * 61000, SDS-PAGE | Plagiochasma appendiculatum |
Synonyms | Comment | Organism |
---|---|---|
flavone synthase I | - |
Plagiochasma appendiculatum |
FNS I | - |
Plagiochasma appendiculatum |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
35 | - |
- |
Plagiochasma appendiculatum |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.002 | - |
naringenin | pH 7.0, 35°C, mutant enzyme Y240P | Plagiochasma appendiculatum | |
0.027 | - |
naringenin | pH 7.0, 35°C, wild-type enzyme | Plagiochasma appendiculatum |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
7 | - |
- |
Plagiochasma appendiculatum |
Organism | Comment | pI Value Maximum | pI Value |
---|---|---|---|
Plagiochasma appendiculatum | calculated from sequence | - |
6.42 |
General Information | Comment | Organism |
---|---|---|
physiological function | involved in flavonoid pathway | Plagiochasma appendiculatum |
kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.305 | - |
naringenin | pH 7.0, 35°C, mutant enzyme Y240P | Plagiochasma appendiculatum | |
1.372 | - |
naringenin | pH 7.0, 35°C, wild-type enzyme | Plagiochasma appendiculatum |