Cloned (Comment) | Organism |
---|---|
gene hcbA3, recombinant expression of C-terminally His-tagged enzyme in Rhodococcus erythropolis, subcloning in Escherichia coli strain DH5alpha | Nocardioides sp. PD653 |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
additional information | - |
additional information | steady-state kinetic analysis, the initial rates of NADH oxidation follow typical Michaelis-Menten kinetics, and Lineweaver-Burk plots show parallel patterns for a ping-pong mechanism | Nocardioides sp. PD653 | |
0.00443 | - |
FMN | recombinant enzyme HcbA3, pH 7.5, 20°C | Nocardioides sp. PD653 | |
0.05166 | - |
NADH | recombinant enzyme HcbA3, pH 7.5, 20°C | Nocardioides sp. PD653 |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
FMN + NADH + H+ | Nocardioides sp. PD653 | - |
FMNH2 + NAD+ | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Nocardioides sp. PD653 | A0A1V1W312 | - |
- |
Purification (Comment) | Organism |
---|---|
recombinant C-terminally His-tagged enzyme from Rhodococcus erythropolis by metal affinity chromatography | Nocardioides sp. PD653 |
Reaction | Comment | Organism | Reaction ID |
---|---|---|---|
FMNH2 + NAD+ = FMN + NADH + H+ | HcbA3 follows a ping-pong bi-bi mechanism for the reduction of flavins | Nocardioides sp. PD653 |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
FMN + NADH + H+ | - |
Nocardioides sp. PD653 | FMNH2 + NAD+ | - |
? | |
additional information | HcbA3 is an NADH:FMN oxidoreductase. Successful reconstitution of the oxidative dehalogenase reaction in vitro, which consists of HcbA1, HcbA3, FMN, and NADH, suggesting that HcbA3 may also be the partner reductase component for HcbA1 in Nocardioides sp. PD653. The optimal molar ratio of HcbA1 and HcbA3 is 7:1 | Nocardioides sp. PD653 | ? | - |
- |
Synonyms | Comment | Organism |
---|---|---|
HcbA3 | - |
Nocardioides sp. PD653 |
hexachlorobenzene oxidative dehalogenase system reductase component | UniProt | Nocardioides sp. PD653 |
NADH:FMN oxidoreductase | - |
Nocardioides sp. PD653 |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
20 | - |
assay at | Nocardioides sp. PD653 |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
66.74 | - |
NADH | recombinant enzyme HcbA3, pH 7.5, 20°C | Nocardioides sp. PD653 | |
66.74 | - |
FMN | recombinant enzyme HcbA3, pH 7.5, 20°C | Nocardioides sp. PD653 |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
7.5 | - |
recombinant His-tagged enzyme | Nocardioides sp. PD653 |
pH Minimum | pH Maximum | Comment | Organism |
---|---|---|---|
7 | 8.6 | at pH 7.0, 7.2, and 8.0, the enzyme activity levels in 100 mM phosphate buffer are approx. 50.2, 88.1, and 81.1%, respectively, relative to that at pH 7.5. HcbA3C-His shows the highest flavin reductase activity in 60 mM KPi buffer | Nocardioides sp. PD653 |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
NADH | - |
Nocardioides sp. PD653 |
General Information | Comment | Organism |
---|---|---|
physiological function | Nocardioides sp. PD653 genes hcbA1, hcbA2, and hcbA3 encode enzymes that catalyze the oxidative dehalogenation of hexachlorobenzene (HCB), which is one of the most recalcitrant persistent organic pollutants (POPs). HcbA3 shows the highest affinity for FMN. HcbA3 may be the partner reductase component for HcbA1 in Nocardioides sp. PD653 | Nocardioides sp. PD653 |
kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
1291.9 | - |
NADH | recombinant enzyme HcbA3, pH 7.5, 20°C | Nocardioides sp. PD653 | |
15065.5 | - |
FMN | recombinant enzyme HcbA3, pH 7.5, 20°C | Nocardioides sp. PD653 |