PT - JOURNAL ARTICLE AU - Jacques W. Bouvier AU - David M. Emms AU - Steven Kelly TI - Slow molecular evolution of rubisco limits adaptive improvement of CO<sub>2</sub> assimilation AID - 10.1101/2022.07.06.498985 DP - 2022 Jan 01 TA - bioRxiv PG - 2022.07.06.498985 4099 - http://biorxiv.org/content/early/2022/07/06/2022.07.06.498985.short 4100 - http://biorxiv.org/content/early/2022/07/06/2022.07.06.498985.full AB - Rubisco assimilates CO2 to form the sugars that fuel life on Earth. Although rubisco is the source of most carbon in the biosphere, it is a surprisingly inefficient catalyst with a modest carboxylase turnover rate and a competing oxygenase activity which results in the loss of fixed CO2. These apparent shortcomings of rubisco present a puzzling evolutionary paradox: why does the enzyme appear well suited to the high CO2 low O2 conditions of its origin, rather than the low CO2 high O2 conditions of the present day? To help answer this question, we perform a phylogenetically resolved analysis of the molecular and kinetic evolution of Form I rubisco. We discover that rubisco is among the slowest evolving genes on Earth. Specifically, we find that the rubisco catalytic large subunit evolves substantially slower than its cognate small subunit, and is slower than &gt;98% of all other genes and enzymes across the tree of life. Next, through simultaneous analysis of rubisco molecular and kinetic evolution in C3 angiosperms, we demonstrate that despite its slow molecular evolution, rubisco kinetics have evolved, and are continuing to evolve, to improve CO2/O2 specificity, carboxylase turnover and overall carboxylation efficiency. Thus, slow molecular evolution has severely limited rubisco kinetic optimisation, resulting in the present enzyme that is poorly adapted for current conditions.Competing Interest StatementThe authors have declared no competing interest.