The activity of RubisCO and energy demands for its biosynthesis. Comparative studies with CO₂-reductases

Most CO₂ on Earth is fixed into organic matter via reactions catalysed by enzymes called carboxylases. CO₂-fixation via carboxylases occurs in the Calvin-Benson-Bassham (CBB) cycle, and the crucial role in this cycle is played by RubisCO (D-ribulose 1,5-bisphosphate carboxylase/oxygenase). CO₂ can also be fixed by pathways, where a reduction of CO₂ to formate or carbon monoxide (CO) occurs. The latter reactions are performed by so-called CO₂-reductases e.g. formate dehydrogenase (FDH), carbon-monooxide (CO) dehydrogenase (CODH), and crotonyl-CoA reductase/carboxylase (CCR). In general, a simple model of enzymatic activity based only on a turnover rate of an enzyme for an appropriate substrate (k_cat) is insufficient. Based on estimated metabolic costs of each amino acid, the average energetic costs of amino acid biosynthesis (E_aa), and the total costs (E_T) for selected CO₂-fixing enzymes were analyzed concerning 1) k_cat for CO₂ (k_C), and 2) specificity factor (S_rel) for RubisCO. A comparison of E_aa and E_T to their k_C showed that CODH and FDHs do not need to be more efficient enzymes in CO₂ capturing pathways than some forms of RubisCO. CCR was the only both low-cost and highly active CO₂-fixing enzyme. The obtained results showed also that there exists an evolutionarily conserved trade-off between S_rel of RubisCOs and the energetic demands needed for their biosynthesis. Phylogenetic analysis demonstrated that RubisCO, CODH, FDH, and CCR are enzymes formed as a result of parallel evolution. Moreover, the kinetic parameters (k_C) of CO₂-fixing enzymes were plausibly optimized already at the early stages of life evolution on Earth.