Modelling the pyrenoid-based CO 2 -concentrating mechanism provides insights into its operating principles and a roadmap for its engineering into crops.

Many eukaryotic photosynthetic organisms enhance their carbon uptake by supplying concentrated CO 2 to the CO 2 -fixing enzyme Rubisco in an organelle called the pyrenoid. Ongoing efforts seek to engineer this pyrenoid-based CO 2 -concentrating mechanism (PCCM) into crops to increase yields. Here we develop a computational model for a PCCM on the basis of the postulated mechanism in the green alga Chlamydomonas reinhardtii. Our model recapitulates all Chlamydomonas PCCM-deficient mutant phenotypes and yields general biophysical principles underlying the PCCM. We show that an effective and energetically efficient PCCM requires a physical barrier to reduce pyrenoid CO 2 leakage, as well as proper enzyme localization to reduce futile cycling between CO 2 and HCO 3 - . Importantly, our model demonstrates the feasibility of a purely passive CO 2 uptake strategy at air-level CO 2 , while active HCO 3 - uptake proves advantageous at lower CO 2 levels. We propose a four-step engineering path to increase the rate of CO 2 fixation in the plant chloroplast up to threefold at a theoretical cost of only 1.3 ATP per CO 2 fixed, thereby offering a framework to guide the engineering of a PCCM into land plants.