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Title: Four amino acids define the CO 2 binding pocket of enoyl-CoA carboxylases/reductases

Abstract

Carboxylases are biocatalysts that capture and convert carbon dioxide (CO 2 ) under mild conditions and atmospheric concentrations at a scale of more than 400 Gt annually. However, how these enzymes bind and control the gaseous CO 2 molecule 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 CO 2 within the active site of Ecr from Kitasatospora 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 CO 2 -fixing enzyme. Together, these 4 residues anchor and position the CO 2 molecule 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 CO 2 fixation. Altogether, our study reveals unprecedented molecular details of selective CO 2 binding and C–C-bond formation during the catalytic cycle of nature’s most efficient COmore » 2 -fixing enzyme. This knowledge provides the basis for the future development of catalytic frameworks for the capture and conversion of CO 2 in biology and chemistry.« less

Authors:
 [1]; ORCiD logo [2];  [3];  [4];  [5]; ORCiD logo [1]; ORCiD logo [6]; ORCiD logo [7];  [2];  [1]
  1. Max Planck Inst. for Terrestrial Microbiology, Marburg (Germany); Center for Synthetic Microbiology, Marburg (Germany)
  2. Univ. de Concepción, Concepción (Chile)
  3. SLAC National Accelerator Lab., Menlo Park, CA (United States); SLAC National Accelerator Lab., Menlo Park, CA (United States). Photon Ultrafast Laser Science and Engineering Inst. (PULSE)
  4. Max Planck Inst. for Terrestrial Microbiology, Marburg (Germany)
  5. SLAC National Accelerator Lab., Menlo Park, CA (United States)
  6. USDOE Joint Genome Institute (JGI), Walnut Creek, CA (United States)
  7. SLAC National Accelerator Lab., Menlo Park, CA (United States); Stanford Univ., CA (United States)
Publication Date:
Research Org.:
SLAC National Accelerator Lab., Menlo Park, CA (United States)
Sponsoring Org.:
USDOE
OSTI Identifier:
1529676
Alternate Identifier(s):
OSTI ID: 1546799
Grant/Contract Number:  
AC02-76SF00515; 637675; AC02-05CH11231; 1231306
Resource Type:
Published Article
Journal Name:
Proceedings of the National Academy of Sciences of the United States of America
Additional Journal Information:
Journal Volume: 116; Journal Issue: 28; Journal ID: ISSN 0027-8424
Publisher:
National Academy of Sciences, Washington, DC (United States)
Country of Publication:
United States
Language:
English
Subject:
37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CHEMISTRY

Citation Formats

Stoffel, Gabriele M. M., Saez, David Adrian, DeMirci, Hasan, Vögeli, Bastian, Rao, Yashas, Zarzycki, Jan, Yoshikuni, Yasuo, Wakatsuki, Soichi, Vöhringer-Martinez, Esteban, and Erb, Tobias J. Four amino acids define the CO2 binding pocket of enoyl-CoA carboxylases/reductases. United States: N. p., 2019. Web. doi:10.1073/pnas.1901471116.
Stoffel, Gabriele M. M., Saez, David Adrian, DeMirci, Hasan, Vögeli, Bastian, Rao, Yashas, Zarzycki, Jan, Yoshikuni, Yasuo, Wakatsuki, Soichi, Vöhringer-Martinez, Esteban, & Erb, Tobias J. Four amino acids define the CO2 binding pocket of enoyl-CoA carboxylases/reductases. United States. doi:10.1073/pnas.1901471116.
Stoffel, Gabriele M. M., Saez, David Adrian, DeMirci, Hasan, Vögeli, Bastian, Rao, Yashas, Zarzycki, Jan, Yoshikuni, Yasuo, Wakatsuki, Soichi, Vöhringer-Martinez, Esteban, and Erb, Tobias J. Wed . "Four amino acids define the CO2 binding pocket of enoyl-CoA carboxylases/reductases". United States. doi:10.1073/pnas.1901471116.
@article{osti_1529676,
title = {Four amino acids define the CO2 binding pocket of enoyl-CoA carboxylases/reductases},
author = {Stoffel, Gabriele M. M. and Saez, David Adrian and DeMirci, Hasan and Vögeli, Bastian and Rao, Yashas and Zarzycki, Jan and Yoshikuni, Yasuo and Wakatsuki, Soichi and Vöhringer-Martinez, Esteban and Erb, Tobias J.},
abstractNote = {Carboxylases are biocatalysts that capture and convert carbon dioxide (CO 2 ) under mild conditions and atmospheric concentrations at a scale of more than 400 Gt annually. However, how these enzymes bind and control the gaseous CO 2 molecule 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 CO 2 within the active site of Ecr from Kitasatospora 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 CO 2 -fixing enzyme. Together, these 4 residues anchor and position the CO 2 molecule 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 CO 2 fixation. Altogether, our study reveals unprecedented molecular details of selective CO 2 binding and C–C-bond formation during the catalytic cycle of nature’s most efficient CO 2 -fixing enzyme. This knowledge provides the basis for the future development of catalytic frameworks for the capture and conversion of CO 2 in biology and chemistry.},
doi = {10.1073/pnas.1901471116},
journal = {Proceedings of the National Academy of Sciences of the United States of America},
number = 28,
volume = 116,
place = {United States},
year = {2019},
month = {6}
}

Journal Article:
Free Publicly Available Full Text
Publisher's Version of Record
DOI: 10.1073/pnas.1901471116

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Works referenced in this record:

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