Cloned (Comment) | Organism |
---|---|
expressed in Escherichia coli DH1 and TG1 | Escherichia coli |
General Stability | Organism |
---|---|
HpaC consists of low stability | Escherichia coli |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.0021 | - |
FMN | - |
Escherichia coli | |
0.0026 | - |
riboflavin | - |
Escherichia coli | |
0.0031 | - |
FAD | - |
Escherichia coli | |
0.04 | - |
NADH | - |
Escherichia coli |
Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
---|---|---|---|
18680 | - |
x * 58781 flavoprotein HpaA, x * 18680, coupling protein, estimation from gene sequence | Klebsiella pneumoniae |
58781 | - |
x * 58781 flavoprotein HpaA, x * 18680, coupling protein, estimation from gene sequence | Klebsiella pneumoniae |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
4-hydroxyphenylacetate + NADH + O2 | Klebsiella pneumoniae | - |
3,4-dihydroxyphenylacetate + NAD+ + H2O | - |
? | |
4-hydroxyphenylacetate + NADH + O2 | Escherichia coli | initial reaction in the degradation of 4-hydroxyphenylacetate | 3,4-dihydroxyphenylacetate + NAD+ + H2O | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Escherichia coli | - |
strain W, ATCC 11105 derived from ATCC 9637 | - |
Klebsiella pneumoniae | - |
- |
- |
Purification (Comment) | Organism |
---|---|
ammonium sulfate precipitation, ion-exchange, gel filtration. Affinity chromatography for the expressed cholin-binding domain containing HpaB protein | Escherichia coli |
Reaction | Comment | Organism | Reaction ID |
---|---|---|---|
4-hydroxyphenylacetate + FADH2 + O2 = 3,4-dihydroxyphenylacetate + FAD + H2O | member of novel two-component flavin-diffusible monooxygenase family, both components required for hydroxylation. The genes are located on the same operon. Physical interaction between HpaB and HpaC is not required, enhancement of activity by direct interaction is not excluded. Contains reductase component | Escherichia coli |
Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
---|---|---|---|
0.14 | - |
activity of HpaB protein in presence of NADH and HpaC protein | Escherichia coli |
0.46 | - |
- |
Escherichia coli |
Storage Stability | Organism |
---|---|
-20°C, HpaC, small protein, very stable, no significant loss of activity during 2 months | Escherichia coli |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
3-hydroxyphenylacetate + FMNH2 + O2 | - |
Escherichia coli | 3,4-hydroxyphenylacetate + FMN + H2O | - |
? | |
4-hydroxyphenylacetate + NADH + O2 | - |
Escherichia coli | 3,4-dihydroxyphenylacetate + NAD+ + H2O | - |
? | |
4-hydroxyphenylacetate + NADH + O2 | - |
Klebsiella pneumoniae | 3,4-dihydroxyphenylacetate + NAD+ + H2O | - |
? | |
4-hydroxyphenylacetate + NADH + O2 | initial reaction in the degradation of 4-hydroxyphenylacetate | Escherichia coli | 3,4-dihydroxyphenylacetate + NAD+ + H2O | - |
? | |
additional information | - |
Klebsiella pneumoniae | ? | - |
? | |
additional information | hydroxylates phenol derivatives | Escherichia coli | ? | - |
? | |
phenol + ? | for HpaB | Escherichia coli | catechol + ? | - |
? | |
riboflavin + NADH + H+ | - |
Escherichia coli | reduced riboflavin + NAD+ | - |
r |
Subunits | Comment | Organism |
---|---|---|
? | - |
Escherichia coli |
? | x * 58781 flavoprotein HpaA, x * 18680, coupling protein, estimation from gene sequence | Klebsiella pneumoniae |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
22 | - |
assay at | Escherichia coli |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
7 | - |
optimal for HpaC protein | Escherichia coli |
7.8 | - |
assay at | Escherichia coli |
pH Minimum | pH Maximum | Comment | Organism |
---|---|---|---|
5 | 8 | more than 80% of activity | Escherichia coli |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
FAD | stimulates | Escherichia coli | |
FAD | stimulates | Klebsiella pneumoniae | |
FMN | used as substrate by HpaC protein | Escherichia coli | |
NADH | - |
Klebsiella pneumoniae | |
NADH | electron donor | Escherichia coli | |
riboflavin | used as substrate by HpaC protein | Escherichia coli |