Four amino acids define the CO 2 binding pocket of enoyl-CoA carboxylases/reductases
[2];
[1];
[6];
[7];
- Department of Biochemistry and Synthetic Metabolism, Max Planck Institute for Terrestrial Microbiology, D-35043 Marburg, Germany,, Center for Synthetic Microbiology, D-35043 Marburg, Germany,
- Departamento de Físico Química, Facultad de Ciencias Químicas, Universidad de Concepción, 4070371 Concepción, Chile,
- Biosciences Division, SLAC National Accelerator Laboratory, Menlo Park, CA 94025,, Stanford PULSE Institute, SLAC National Accelerator Laboratory, Menlo Park, CA 94025,
- Department of Biochemistry and Synthetic Metabolism, Max Planck Institute for Terrestrial Microbiology, D-35043 Marburg, Germany,
- Biosciences Division, SLAC National Accelerator Laboratory, Menlo Park, CA 94025,
- Department of Energy Joint Genome Institute, Walnut Creek, CA 94598,
- Biosciences Division, SLAC National Accelerator Laboratory, Menlo Park, CA 94025,, Structural Biology Department, Stanford University, Stanford, CA 94305
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.
- Research Organization:
- SLAC National Accelerator Laboratory (SLAC), Menlo Park, CA (United States); Lawrence Berkeley National Laboratory (LBNL), Berkeley, CA (United States)
- Sponsoring Organization:
- USDOE Office of Science (SC), Biological and Environmental Research (BER)
- Grant/Contract Number:
- AC02-05CH11231; AC02-76SF00515; 637675; 1231306
- OSTI ID:
- 1529676
- Alternate ID(s):
- OSTI ID: 1546799; OSTI ID: 1559229
- Journal Information:
- Proceedings of the National Academy of Sciences of the United States of America, Journal Name: Proceedings of the National Academy of Sciences of the United States of America Vol. 116 Journal Issue: 28; ISSN 0027-8424
- Publisher:
- Proceedings of the National Academy of SciencesCopyright Statement
- Country of Publication:
- United States
- Language:
- English
Web of Science
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