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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:
; ORCiD logo; ; ; ; ORCiD logo; ORCiD logo; ORCiD logo; ;
Publication Date:
Sponsoring Org.:
USDOE
OSTI Identifier:
1529676
Grant/Contract Number:  
AC02-05CH11231
Resource Type:
Published Article
Journal Name:
Proceedings of the National Academy of Sciences of the United States of America
Additional Journal Information:
Journal Name: Proceedings of the National Academy of Sciences of the United States of America Journal Volume: 116 Journal Issue: 28; Journal ID: ISSN 0027-8424
Publisher:
Proceedings of the National Academy of Sciences
Country of Publication:
United States
Language:
English

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 CO 2 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 CO 2 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 CO 2 binding pocket of enoyl-CoA carboxylases/reductases". United States. doi:10.1073/pnas.1901471116.
@article{osti_1529676,
title = {Four amino acids define the CO 2 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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