Cloned (Comment) | Organism |
---|---|
expressed in Escherichia coli as a His-tagged fusion protein | Synechococcus |
Protein Variants | Comment | Organism |
---|---|---|
A375I | mutant shows only insignificant activity and is not able to complement Rhodobacter capsulatus strain SBI/II- (Rubisco knockout strain) | Synechococcus |
A375S | mutant shows only insignificant activity and is not able to complement Rhodobacter capsulatus strain SBI/II- (Rubisco knockout strain) | Synechococcus |
A375V | kcat is highly reduced compared to wild-type, A375V shows 12% activity compared to wild-type, mutant A375V is fully capable of supporting anaerobic phototrophic CO2-dependent growth of the Rhodobacter capsulatus strain SBI/II- (Rubisco knockout strain). Unlike the wild-type Synechococcus enzyme, the A375V mutant is able to support anaerobic photoautotrophic growth at a limiting (1.5%) CO2 concentration. Mutant A375V is able to support aerobic CO2-dependent chemoautotrophic growth and enhance aerobic mixotrophic growth in a complex organic growth media. In contrast to wild-type mutant A375V maintains about 60% of its original activity after a 10 min incubation at 65°C. Thus, the A375V substitution has a pronounced effect on both the structure and function of the enzyme. Km (CO2) value is 23% lower compared to wild-type. Km (O2) value is elevated compared to wild-type. Km (D-ribulose 1,5-bisphosphate) moderately elevated compared to wild-type | Synechococcus |
D103V | mutant fails to support CO2-dependent growth of cultures of Rhodobacter capsulatus since surface residues involved with large subunit interaction influence the Michaelis constant for CO2, kcat is equal to wild-type, mutant D103V shows comparable amounts of Rubisco activity to wild-type after a 10 min incubation at 65°C. Km (CO2) value higher compared to wild-type. Km (O2) and (D-ribulose 1,5-bisphosphate) value equal to wild-type | Synechococcus |
D103V/A375V | kcat is moderately reduced compared to wild-type, mutant D103V/A375V shows 63% activity compared to wild-type,mutant A375V is fully capable of supporting anaerobic phototrophic CO2-dependent growth of the Rhodobacter capsulatus strain SBI/II- (Rubisco knockout strain). The A375V substitution on the same gene as the D103V substitution is an intragenic suppressor that restores photoautotrophic growth when the resulting enzyme is used to complement the Rhodobacter capsulatus strain SBI/II- (Rubisco knockout strain). DOuble mutnat D103V/A375V is to a lesser extent (compared to A375V) able to support aerobic CO2-dependent chemoautotrophic growth and enhance aerobic mixotrophic growth in a complex organic growth media. Mutant D103V/A375V shows comparable amounts of Rubisco activity to wild-type after a 10 min incubation at 65°C. Km (CO2) is moderately elevated compared to wild-type. Km (O2) and (D-ribulose 1,5-bisphosphate) elevated compared to wild-type | Synechococcus |
F97L | mutant is not able to complement Rhodobacter capsulatus strain SBI/II- (Rubisco knockout strain) | Synechococcus |
F97L/D103V | mutant is not able to complement Rhodobacter capsulatus strain SBI/II- (Rubisco knockout strain) | Synechococcus |
G176D | mutant is not able to complement Rhodobacter capsulatus strain SBI/II- (Rubisco knockout strain) | Synechococcus |
G176D/A375V | mutant is not able to complement Rhodobacter capsulatus strain SBI/II- (Rubisco knockout strain). A375V substitution is unable to suppress the negative phenotype of G176D | Synechococcus |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.025 | - |
D-ribulose 1,5-bisphosphate | mutant D103V | Synechococcus | |
0.027 | - |
D-ribulose 1,5-bisphosphate | wild-type | Synechococcus | |
0.034 | - |
D-ribulose 1,5-bisphosphate | mutant A375V | Synechococcus | |
0.143 | - |
CO2 | mutant A375V | Synechococcus | |
0.186 | - |
CO2 | wild-type | Synechococcus | |
0.269 | - |
D-ribulose 1,5-bisphosphate | mutant D103V/A375V | Synechococcus | |
0.294 | - |
CO2 | mutant D103V/A375V | Synechococcus | |
0.388 | - |
CO2 | mutant D103V | Synechococcus | |
0.85 | - |
O2 | wild-type | Synechococcus | |
0.866 | - |
O2 | mutant D103V | Synechococcus | |
1.088 | - |
O2 | mutant A375V | Synechococcus | |
1.153 | - |
O2 | mutant D103V/A375V | Synechococcus |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Synechococcus | - |
- |
- |
Purification (Comment) | Organism |
---|---|
using Ni-NTA chromatography and ion-exchange chromatography | Synechococcus |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
D-Ribulose 1,5-bisphosphate + CO2 | - |
Synechococcus | ? | - |
? | |
D-ribulose 1,5-bisphosphate + O2 + H2O | - |
Synechococcus | ? | - |
? |
Synonyms | Comment | Organism |
---|---|---|
Ribulose 1,5-bisphosphate carboxylase/oxygenase | - |
Synechococcus |
Rubisco | - |
Synechococcus |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.6 | - |
D-ribulose 1,5-bisphosphate | mutant A375V, co-substrate: CO2 | Synechococcus | |
0.6 | - |
CO2 | mutant A375V, co-substrate: D-ribulose 1,5-bisphosphate | Synechococcus | |
3.1 | - |
CO2 | mutant A375V/D103V, co-substrate: D-ribulose 1,5-bisphosphate | Synechococcus | |
3.1 | - |
D-ribulose 1,5-bisphosphate | mutant D103V/A375V, co-substrate: CO2 | Synechococcus | |
4.9 | - |
D-ribulose 1,5-bisphosphate | mutant D103V, co-substrate: CO2 | Synechococcus | |
4.9 | - |
CO2 | mutant D103V, co-substrate: D-ribulose 1,5-bisphosphate | Synechococcus | |
4.9 | - |
D-ribulose 1,5-bisphosphate | wild-type, co-substrate: CO2 | Synechococcus | |
4.9 | - |
CO2 | wild-type, co-substrate: D-ribulose 1,5-bisphosphate | Synechococcus |