Noam Prywes, Naiya R Philips, Luke M Oltrogge, Sebastian Lindner, Yi-Chin Candace Tsai, Benoit de Pins, Aidan E Cowan, Leah J Taylor-Kearney, Hana A Chang, Laina N Hall, Daniel Bellieny-Rabelo, Hunter M Nisonoff, Rachel F Weissman, Avi I Flamholz, David Ding, Abhishek Y Bhatt, Patrick M Shih, Oliver Mueller-Cajar, Ron Milo, David F Savage
Rubisco is the primary CO 2 fixing enzyme of the biosphere yet has slow kinetics. The roles of evolution and chemical mechanism in constraining the sequence landscape of rubisco remain debated. In order to map sequence to function, we developed a massively parallel assay for rubisco using an engineered E. coli where enzyme function is coupled to growth. By assaying >99% of single amino acid mutants across CO 2 concentrations, we inferred enzyme velocity and CO 2 affinity for thousands of substitutions. We identified many highly conserved positions that tolerate mutation and rare mutations that improve CO 2 affinity...
April 11, 2024: bioRxiv