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Title: Reaction kinetic analysis of the 3-hydroxypropionate/4-hydroxybutyrate CO 2 fixation cycle in extremely thermoacidophilic archaea

Abstract

Here, the 3-hydroxypropionate/4-hydroxybutyrate (3HP/4HB) cycle fixes CO 2 in extremely thermoacidophilic archaea and holds promise for metabolic engineering because of its thermostability and potentially rapid pathway kinetics. A reaction kinetics model was developed to examine the biological and biotechnological attributes of the 3HP/4HB cycle as it operates in Metallosphaera sedula, based on previous information as well as on kinetic parameters determined here for recombinant versions of five of the cycle enzymes (malonyl-CoA/succinyl-CoA reductase, 3-hydroxypropionyl-CoA synthetase, 3-hydroxypropionyl-CoA dehydratase, acryloyl-CoA reductase, and succinic semialdehyde reductase). The model correctly predicted previously observed features of the cycle: the 35%–65% split of carbon flux through the acetyl-CoA and succinate branches, the high abundance and relative ratio of acetyl-CoA/propionyl-CoA carboxylase (ACC) and MCR, and the significance of ACC and hydroxybutyryl-CoA synthetase (HBCS) as regulated control points for the cycle. The model was then used to assess metabolic engineering strategies for incorporating CO 2 into chemical intermediates and products of biotechnological importance: acetyl-CoA, succinate, and 3-hydroxyproprionate.

Authors:
 [1];  [2];  [1];  [3];  [4];  [4];  [4];  [4];  [5]
  1. North Carolina State Univ., Raleigh, NC (United States); Novozymes North America Inc., Franklinton, NC (United States)
  2. North Carolina State Univ., Raleigh, NC (United States); Chinese Academy of Sciences, Beijing (China)
  3. North Carolina State Univ., Raleigh, NC (United States); Xiamen Univ., Fujan Province (China)
  4. Univ. of Georgia, Athens, GA (United States)
  5. North Carolina State Univ., Raleigh, NC (United States)
Publication Date:
Research Org.:
North Carolina State Univ., Raleigh, NC (United States)
Sponsoring Org.:
USDOE Advanced Research Projects Agency - Energy (ARPA-E)
OSTI Identifier:
1422400
Alternate Identifier(s):
OSTI ID: 1410825
Grant/Contract Number:  
AR0000081
Resource Type:
Journal Article: Accepted Manuscript
Journal Name:
Metabolic Engineering
Additional Journal Information:
Journal Volume: 38; Journal Issue: C; Journal ID: ISSN 1096-7176
Publisher:
Elsevier
Country of Publication:
United States
Language:
English
Subject:
37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CHEMISTRY; CO2 fixation; 3-hydroxypropionate; 4-hydroxybutyrate; Metallosphaera sedula

Citation Formats

Loder, Andrew J., Han, Yejun, Hawkins, Aaron B., Lian, Hong, Lipscomb, Gina L., Schut, Gerrit J., Keller, Matthew W., Adams, Michael W. W., and Kelly, Robert M. Reaction kinetic analysis of the 3-hydroxypropionate/4-hydroxybutyrate CO2 fixation cycle in extremely thermoacidophilic archaea. United States: N. p., 2016. Web. doi:10.1016/j.ymben.2016.10.009.
Loder, Andrew J., Han, Yejun, Hawkins, Aaron B., Lian, Hong, Lipscomb, Gina L., Schut, Gerrit J., Keller, Matthew W., Adams, Michael W. W., & Kelly, Robert M. Reaction kinetic analysis of the 3-hydroxypropionate/4-hydroxybutyrate CO2 fixation cycle in extremely thermoacidophilic archaea. United States. doi:10.1016/j.ymben.2016.10.009.
Loder, Andrew J., Han, Yejun, Hawkins, Aaron B., Lian, Hong, Lipscomb, Gina L., Schut, Gerrit J., Keller, Matthew W., Adams, Michael W. W., and Kelly, Robert M. Wed . "Reaction kinetic analysis of the 3-hydroxypropionate/4-hydroxybutyrate CO2 fixation cycle in extremely thermoacidophilic archaea". United States. doi:10.1016/j.ymben.2016.10.009. https://www.osti.gov/servlets/purl/1422400.
@article{osti_1422400,
title = {Reaction kinetic analysis of the 3-hydroxypropionate/4-hydroxybutyrate CO2 fixation cycle in extremely thermoacidophilic archaea},
author = {Loder, Andrew J. and Han, Yejun and Hawkins, Aaron B. and Lian, Hong and Lipscomb, Gina L. and Schut, Gerrit J. and Keller, Matthew W. and Adams, Michael W. W. and Kelly, Robert M.},
abstractNote = {Here, the 3-hydroxypropionate/4-hydroxybutyrate (3HP/4HB) cycle fixes CO2 in extremely thermoacidophilic archaea and holds promise for metabolic engineering because of its thermostability and potentially rapid pathway kinetics. A reaction kinetics model was developed to examine the biological and biotechnological attributes of the 3HP/4HB cycle as it operates in Metallosphaera sedula, based on previous information as well as on kinetic parameters determined here for recombinant versions of five of the cycle enzymes (malonyl-CoA/succinyl-CoA reductase, 3-hydroxypropionyl-CoA synthetase, 3-hydroxypropionyl-CoA dehydratase, acryloyl-CoA reductase, and succinic semialdehyde reductase). The model correctly predicted previously observed features of the cycle: the 35%–65% split of carbon flux through the acetyl-CoA and succinate branches, the high abundance and relative ratio of acetyl-CoA/propionyl-CoA carboxylase (ACC) and MCR, and the significance of ACC and hydroxybutyryl-CoA synthetase (HBCS) as regulated control points for the cycle. The model was then used to assess metabolic engineering strategies for incorporating CO2 into chemical intermediates and products of biotechnological importance: acetyl-CoA, succinate, and 3-hydroxyproprionate.},
doi = {10.1016/j.ymben.2016.10.009},
journal = {Metabolic Engineering},
issn = {1096-7176},
number = C,
volume = 38,
place = {United States},
year = {2016},
month = {10}
}

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