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Literature summary for 4.1.1.39 extracted from

  • Satagopan, S.; Scott, S.S.; Smith, T.G.; Tabita, F.R.
    A Rubisco mutant that confers growth under a normally "inhibitory" oxygen concentration (2009), Biochemistry, 48, 9076-9083.
    View publication on PubMedView publication on EuropePMC

Cloned(Commentary)

Cloned (Comment) Organism
expressed in Escherichia coli as a His-tagged fusion protein Synechococcus

Protein Variants

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 [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

Organism UniProt Comment Textmining
Synechococcus
-
-
-

Purification (Commentary)

Purification (Comment) Organism
using Ni-NTA chromatography and ion-exchange chromatography Synechococcus

Substrates and Products (Substrate)

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

Synonyms Comment Organism
Ribulose 1,5-bisphosphate carboxylase/oxygenase
-
Synechococcus
Rubisco
-
Synechococcus

Turnover Number [1/s]

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