This paper presents the development of glycolyl-CoA carboxylase (GCC), a novel enzyme engineered to enhance CO2 fixation. Through rational design, high-throughput microfluidics, and microplate screens, GCC's catalytic efficiency was improved by three orders of magnitude. The redesigned active site was validated using cryo-electron microscopy. A carboxylation module, including GCC, was created for converting glycolate (C2) to glycerate (C3), and its application in photorespiration, ethylene glycol conversion, and synthetic CO2 fixation was demonstrated. Stoichiometric calculations suggest GCC could increase carbon efficiency by up to 150% and reduce energy demand.

Marieke Scheffen, Daniel G. Marchal, Thomas Beneyton, Sandra K. Schuller, Melanie Klose, Christoph Diehl, Jessica Lehmann, Pascal Pfister, Martina Carrillo, Hai He, Selçuk Aslan, Niña S. Cortina, Peter Claus, Daniel Bollschweiler, Jean-Christophe Baret, Jan M. Schuller, Jan Zarzycki, Arren Bar-Even, Tobias J. Erb