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Gabriele M. M. Stoffel and David Adrian Saez and Hasan DeMirci and Bastian Vögeli and Yashas Rao and Jan Zarzycki and Yasuo Yoshikuni and Soichi Wakatsuki and Esteban Vöhringer-Martinez and Tobias J. Erb

Stoffel, G.M.M. et al., 2019. Four amino acids define the CO2binding pocket of enoyl-CoA carboxylases/reductases. Proceedings of the National Academy of Sciences, 116(28), pp.13964–13969. Available at: http://dx.doi.org/10.1073/pnas.1901471116.

Carboxylases are biocatalysts that capture and convert carbon dioxide (CO2) under mild conditions and atmospheric concentrations at a scale of more than 400 Gt annually. However, how these enzymes bind and control the gaseous CO2molecule during catalysis is only poorly understood. One of the most efficient classes of carboxylating enzymes are enoyl-CoA carboxylases/reductases (Ecrs), which outcompete the plant enzyme RuBisCO in catalytic efficiency and fidelity by more than an order of magnitude. Here we investigated the interactions of CO2within the active site of Ecr fromKitasatospora setae. Combining experimental biochemistry, protein crystallography, and advanced computer simulations we show that 4 amino acids, N81, F170, E171, and H365, are required to create a highly efficient CO2-fixing enzyme. Together, these 4 residues anchor and position the CO2molecule for the attack by a reactive enolate created during the catalytic cycle. Notably, a highly ordered water molecule plays an important role in an active site that is otherwise carefully shielded from water, which is detrimental to CO2fixation. Altogether, our study reveals unprecedented molecular details of selective CO2binding and C–C-bond formation during the catalytic cycle of nature’s most efficient CO2-fixing enzyme. This knowledge provides the basis for the future development of catalytic frameworks for the capture and conversion of CO2in biology and chemistry.

http://dx.doi.org/10.1073/pnas.1901471116

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