DOE PAGES title logo U.S. Department of Energy
Office of Scientific and Technical Information

Title: Augmenting the Calvin–Benson–Bassham cycle by a synthetic malyl-CoA-glycerate carbon fixation pathway

Abstract

The Calvin–Benson–Bassham (CBB) cycle is presumably evolved for optimal synthesis of C3 sugars, but not for the production of C2 metabolite acetyl-CoA. The carbon loss in producing acetyl-CoA from decarboxylation of C3 sugar limits the maximum carbon yield of photosynthesis. Here we design a synthetic malyl-CoA-glycerate (MCG) pathway to augment the CBB cycle for efficient acetyl-CoA synthesis. This pathway converts a C3 metabolite to two acetyl-CoA by fixation of one additional CO2 equivalent, or assimilates glyoxylate, a photorespiration intermediate, to produce acetyl-CoA without net carbon loss. We first functionally demonstrate the design of the MCG pathway in vitro and in Escherichia coli. We then implement the pathway in a photosynthetic organism Synechococcus elongates PCC7942, and show that it increases the intracellular acetyl-CoA pool and enhances bicarbonate assimilation by roughly 2-fold. This work provides a strategy to improve carbon fixation efficiency in photosynthetic organisms.

Authors:
 [1];  [1];  [1];  [1];  [2]
  1. Univ. of California, Los Angeles, CA (United States)
  2. Academia Sinica, Taipei (Taiwan)
Publication Date:
Research Org.:
Univ. of California, Los Angeles, CA (United States)
Sponsoring Org.:
USDOE Advanced Research Projects Agency - Energy (ARPA-E)
OSTI Identifier:
1500021
Grant/Contract Number:  
AR0000201; FC03-02ER63421
Resource Type:
Accepted Manuscript
Journal Name:
Nature Communications
Additional Journal Information:
Journal Volume: 9; Journal Issue: 1; Journal ID: ISSN 2041-1723
Publisher:
Nature Publishing Group
Country of Publication:
United States
Language:
English
Subject:
59 BASIC BIOLOGICAL SCIENCES

Citation Formats

Yu, Hong, Li, Xiaoqian, Duchoud, Fabienne, Chuang, Derrick S., and Liao, James C. Augmenting the Calvin–Benson–Bassham cycle by a synthetic malyl-CoA-glycerate carbon fixation pathway. United States: N. p., 2018. Web. doi:10.1038/s41467-018-04417-z.
Yu, Hong, Li, Xiaoqian, Duchoud, Fabienne, Chuang, Derrick S., & Liao, James C. Augmenting the Calvin–Benson–Bassham cycle by a synthetic malyl-CoA-glycerate carbon fixation pathway. United States. https://doi.org/10.1038/s41467-018-04417-z
Yu, Hong, Li, Xiaoqian, Duchoud, Fabienne, Chuang, Derrick S., and Liao, James C. Tue . "Augmenting the Calvin–Benson–Bassham cycle by a synthetic malyl-CoA-glycerate carbon fixation pathway". United States. https://doi.org/10.1038/s41467-018-04417-z. https://www.osti.gov/servlets/purl/1500021.
@article{osti_1500021,
title = {Augmenting the Calvin–Benson–Bassham cycle by a synthetic malyl-CoA-glycerate carbon fixation pathway},
author = {Yu, Hong and Li, Xiaoqian and Duchoud, Fabienne and Chuang, Derrick S. and Liao, James C.},
abstractNote = {The Calvin–Benson–Bassham (CBB) cycle is presumably evolved for optimal synthesis of C3 sugars, but not for the production of C2 metabolite acetyl-CoA. The carbon loss in producing acetyl-CoA from decarboxylation of C3 sugar limits the maximum carbon yield of photosynthesis. Here we design a synthetic malyl-CoA-glycerate (MCG) pathway to augment the CBB cycle for efficient acetyl-CoA synthesis. This pathway converts a C3 metabolite to two acetyl-CoA by fixation of one additional CO2 equivalent, or assimilates glyoxylate, a photorespiration intermediate, to produce acetyl-CoA without net carbon loss. We first functionally demonstrate the design of the MCG pathway in vitro and in Escherichia coli. We then implement the pathway in a photosynthetic organism Synechococcus elongates PCC7942, and show that it increases the intracellular acetyl-CoA pool and enhances bicarbonate assimilation by roughly 2-fold. This work provides a strategy to improve carbon fixation efficiency in photosynthetic organisms.},
doi = {10.1038/s41467-018-04417-z},
journal = {Nature Communications},
number = 1,
volume = 9,
place = {United States},
year = {Tue May 22 00:00:00 EDT 2018},
month = {Tue May 22 00:00:00 EDT 2018}
}

Journal Article:
Free Publicly Available Full Text
Publisher's Version of Record

Citation Metrics:
Cited by: 60 works
Citation information provided by
Web of Science

Figures / Tables:

Table 1 Table 1: Comparison of different pathway combinations for synthesizing each acetyl-CoA from CO2 equivalents

Save / Share:

Works referenced in this record:

Characterization of the D-Xylulose 5-Phosphate/D-Fructose 6-Phosphate Phosphoketolase Gene (xfp) from Bifidobacterium lactis
journal, May 2001


Prospects for improving CO2 fixation in C3-crops through understanding C4-Rubisco biogenesis and catalytic diversity
journal, June 2016

  • Sharwood, Robert E.; Ghannoum, Oula; Whitney, Spencer M.
  • Current Opinion in Plant Biology, Vol. 31
  • DOI: 10.1016/j.pbi.2016.04.002

pH-Dependent regulation of electron transport and ATP synthesis in chloroplasts
journal, May 2013


Structure of the Escherichia coli Fumarate Reductase Respiratory Complex
journal, June 1999


Structure of the Escherichia coli Fumarate Reductase Respiratory Complex
journal, June 1999


Microbial synthesis of n-butanol, isobutanol, and other higher alcohols from diverse resources
journal, May 2013


Acetate and Bicarbonate Assimilation and Metabolite Formation in Chlamydomonas reinhardtii: A 13C-NMR Study
journal, September 2014


Stereochemical course of reactions catalyzed by the bacterial phosphoenolpyruvate: glucose phosphotransferase system
journal, October 1982

  • Begley, Gail S.; Hansen, David E.; Jacobson, Gary R.
  • Biochemistry, Vol. 21, Issue 22
  • DOI: 10.1021/bi00265a026

An Escherichia coli mutant deficient in pyruvate oxidase activity due to altered phospholipid activation of the enzyme
journal, July 1984

  • Chang, Y. -Y.; Cronan, J. E.
  • Proceedings of the National Academy of Sciences, Vol. 81, Issue 14
  • DOI: 10.1073/pnas.81.14.4348

Strategies for improving C4 photosynthesis
journal, June 2016


A faster Rubisco with potential to increase photosynthesis in crops
journal, September 2014

  • Lin, Myat T.; Occhialini, Alessandro; Andralojc, P. John
  • Nature, Vol. 513, Issue 7519
  • DOI: 10.1038/nature13776

Photorespiration and the potential to improve photosynthesis
journal, December 2016


Metabolic engineering of Escherichia coli for 1-butanol production
journal, November 2008

  • Atsumi, Shota; Cann, Anthony F.; Connor, Michael R.
  • Metabolic Engineering, Vol. 10, Issue 6, p. 305-311
  • DOI: 10.1016/j.ymben.2007.08.003

Enhancing C3 photosynthesis: an outlook on feasible interventions for crop improvement
journal, September 2014

  • Singh, Jitender; Pandey, Prachi; James, Donald
  • Plant Biotechnology Journal, Vol. 12, Issue 9
  • DOI: 10.1111/pbi.12246

A faster Rubisco with potential to increase photosynthesis in crops
journal, September 2014

  • Lin, Myat T.; Occhialini, Alessandro; Andralojc, P. John
  • Nature, Vol. 513, Issue 7519
  • DOI: 10.1038/nature13776

High-performance liquid chromatographic determination of glyoxylic acid in urine
journal, January 1988

  • Petrarulo, Michele; Pellegrino, Sergio; Bianco, Ornella
  • Journal of Chromatography B: Biomedical Sciences and Applications, Vol. 432
  • DOI: 10.1016/S0378-4347(00)80631-3

Distribution of the Isopropylmalate Pathway to Leucine Among Diverse Bacteria
journal, January 1974


Engineering fatty acid biosynthesis in microalgae for sustainable biodiesel
journal, June 2013

  • Blatti, Jillian L.; Michaud, Jennifer; Burkart, Michael D.
  • Current Opinion in Chemical Biology, Vol. 17, Issue 3
  • DOI: 10.1016/j.cbpa.2013.04.007

Molecular Characterization of Escherichia coli Malate Synthase G. Differentiation with the Malate Synthase A Isoenzyme
journal, September 1994


Cross-induction of glc and ace Operons of Escherichia coli Attributable to Pathway Intersection : CHARACTERIZATION OF THE
journal, January 1999

  • Pellicer, Maria Teresa; Fernandez, Carmen; Badı́a, Josefa
  • Journal of Biological Chemistry, Vol. 274, Issue 3
  • DOI: 10.1074/jbc.274.3.1745

Direct photosynthetic recycling of carbon dioxide to isobutyraldehyde
journal, November 2009

  • Atsumi, Shota; Higashide, Wendy; Liao, James C.
  • Nature Biotechnology, Vol. 27, Issue 12, p. 1177-1180
  • DOI: 10.1038/nbt.1586

Amino Acid Biosynthesis and its Regulation
journal, June 1978


Enzymatic assembly of DNA molecules up to several hundred kilobases
journal, April 2009

  • Gibson, Daniel G.; Young, Lei; Chuang, Ray-Yuan
  • Nature Methods, Vol. 6, Issue 5, p. 343-345
  • DOI: 10.1038/nmeth.1318

Engineering Cyanobacteria To Synthesize and Export Hydrophilic Products
journal, April 2010

  • Niederholtmeyer, H.; Wolfstadter, B. T.; Savage, D. F.
  • Applied and Environmental Microbiology, Vol. 76, Issue 11
  • DOI: 10.1128/AEM.00202-10

Non-fermentative pathways for synthesis of branched-chain higher alcohols as biofuels
journal, January 2008

  • Atsumi, Shota; Hanai, Taizo; Liao, James C.
  • Nature, Vol. 451, Issue 7174, p. 86-89
  • DOI: 10.1038/nature06450

A synthetic pathway for the fixation of carbon dioxide in vitro
journal, November 2016

  • Schwander, Thomas; Schada von Borzyskowski, Lennart; Burgener, Simon
  • Science, Vol. 354, Issue 6314
  • DOI: 10.1126/science.aah5237

Rubisco without the Calvin cycle improves the carbon efficiency of developing green seeds
journal, December 2004

  • Schwender, Jörg; Goffman, Fernando; Ohlrogge, John B.
  • Nature, Vol. 432, Issue 7018
  • DOI: 10.1038/nature03145

Acetate and Bicarbonate Assimilation and Metabolite Formation in Chlamydomonas reinhardtii: A 13C-NMR Study
journal, September 2014


A reverse glyoxylate shunt to build a non-native route from C4 to C2 in Escherichia coli
journal, September 2013


Ensemble Modeling for Robustness Analysis in engineering non-native metabolic pathways
journal, September 2014


A critical review on the improvement of photosynthetic carbon assimilation in C 3 plants using genetic engineering
journal, June 2011

  • Ruan, Cheng-Jiang; Shao, Hong-Bo; Teixeira da Silva, Jaime A.
  • Critical Reviews in Biotechnology, Vol. 32, Issue 1
  • DOI: 10.3109/07388551.2010.533119

A reverse glyoxylate shunt to build a non-native route from C4 to C2 in Escherichia coli
journal, September 2013


A radical-chemical route to acetyl-CoA: the anaerobically induced pyruvate formate-lyase system of Escherichia coli
journal, August 1990


Rubisco without the Calvin cycle improves the carbon efficiency of developing green seeds
journal, December 2004

  • Schwender, Jörg; Goffman, Fernando; Ohlrogge, John B.
  • Nature, Vol. 432, Issue 7018
  • DOI: 10.1038/nature03145

Current methods for estimating the rate of photorespiration in leaves
journal, November 2012


Ensemble Modeling for Robustness Analysis in engineering non-native metabolic pathways
journal, September 2014


Overexpression of SBPase enhances photosynthesis against high temperature stress in transgenic rice plants
journal, April 2007


Metabolic Engineering of Escherichia coli for Production of Mixed-Acid Fermentation End Products
journal, May 2014

  • Förster, Andreas H.; Gescher, Johannes
  • Frontiers in Bioengineering and Biotechnology, Vol. 2
  • DOI: 10.3389/fbioe.2014.00016

Chloroplastic photorespiratory bypass increases photosynthesis and biomass production in Arabidopsis thaliana
journal, April 2007

  • Kebeish, Rashad; Niessen, Markus; Thiruveedhi, Krishnaveni
  • Nature Biotechnology, Vol. 25, Issue 5
  • DOI: 10.1038/nbt1299

Chloroplastic photorespiratory bypass increases photosynthesis and biomass production in Arabidopsis thaliana
journal, April 2007

  • Kebeish, Rashad; Niessen, Markus; Thiruveedhi, Krishnaveni
  • Nature Biotechnology, Vol. 25, Issue 5
  • DOI: 10.1038/nbt1299

Synthetic non-oxidative glycolysis enables complete carbon conservation
journal, September 2013

  • Bogorad, Igor W.; Lin, Tzu-Shyang; Liao, James C.
  • Nature, Vol. 502, Issue 7473
  • DOI: 10.1038/nature12575

Over-expression, purification, and characterization of recombinant NAD-malic enzyme from Escherichia coli K12
journal, May 2007

  • Wang, Jinxia; Tan, Haidong; Zhao, Zongbao (Kent)
  • Protein Expression and Purification, Vol. 53, Issue 1
  • DOI: 10.1016/j.pep.2006.11.017

Building carbon–carbon bonds using a biocatalytic methanol condensation cycle
journal, October 2014

  • Bogorad, Igor W.; Chen, Chang-Ting; Theisen, Matthew K.
  • Proceedings of the National Academy of Sciences, Vol. 111, Issue 45
  • DOI: 10.1073/pnas.1413470111

Recent progress in the metabolic engineering of alkaloids in plant systems
journal, April 2013

  • Glenn, Weslee S.; Runguphan, Weerawat; O’Connor, Sarah E.
  • Current Opinion in Biotechnology, Vol. 24, Issue 2
  • DOI: 10.1016/j.copbio.2012.08.003

Overexpression of SBPase enhances photosynthesis against high temperature stress in transgenic rice plants
journal, April 2007


Machine learning discovery of missing links that mediate alternative branches to plant alkaloids
journal, March 2022

  • Vavricka, Christopher J.; Takahashi, Shunsuke; Watanabe, Naoki
  • Nature Communications, Vol. 13, Issue 1
  • DOI: 10.1038/s41467-022-28883-8

Escherichia coli Malic Enzymes: Two Isoforms with Substantial Differences in Kinetic Properties, Metabolic Regulation, and Structure
journal, June 2007

  • Bologna, F. P.; Andreo, C. S.; Drincovich, M. F.
  • Journal of Bacteriology, Vol. 189, Issue 16
  • DOI: 10.1128/jb.00428-07

Current methods for estimating the rate of photorespiration in leaves
journal, November 2012


The IdhA Gene Encoding the Fermentative Lactate Dehydrogenase of Escherichia Coli
journal, January 1997


Design and analysis of synthetic carbon fixation pathways
journal, April 2010

  • Bar-Even, A.; Noor, E.; Lewis, N. E.
  • Proceedings of the National Academy of Sciences, Vol. 107, Issue 19, p. 8889-8894
  • DOI: 10.1073/pnas.0907176107

Ammonia-oxidizing archaea use the most energy-efficient aerobic pathway for CO2 fixation
journal, May 2014

  • Konneke, M.; Schubert, D. M.; Brown, P. C.
  • Proceedings of the National Academy of Sciences, Vol. 111, Issue 22
  • DOI: 10.1073/pnas.1402028111

Metabolic engineering of Escherichia coli for 1-butanol production
journal, November 2008

  • Atsumi, Shota; Cann, Anthony F.; Connor, Michael R.
  • Metabolic Engineering, Vol. 10, Issue 6, p. 305-311
  • DOI: 10.1016/j.ymben.2007.08.003

Enhancing C3 photosynthesis: an outlook on feasible interventions for crop improvement
journal, September 2014

  • Singh, Jitender; Pandey, Prachi; James, Donald
  • Plant Biotechnology Journal, Vol. 12, Issue 9
  • DOI: 10.1111/pbi.12246

Structure, Expression, and Protein Engineering of the Pyruvate Dehydrogenase Complex of Escherichia coli
journal, December 1989


Energy and carbon accounting to compare bioenergy crops
journal, June 2013


Comparison of individual component deletions in a glucose-specific phosphotransferase system revealed their different applications
journal, August 2015

  • Liang, Quanfeng; Zhang, Fengyu; Li, Yikui
  • Scientific Reports, Vol. 5, Issue 1
  • DOI: 10.1038/srep13200

Photorespiration and the potential to improve photosynthesis
journal, December 2016


A synthetic pathway for the fixation of carbon dioxide in vitro
journal, November 2016

  • Schwander, Thomas; Schada von Borzyskowski, Lennart; Burgener, Simon
  • Science, Vol. 354, Issue 6314
  • DOI: 10.1126/science.aah5237

Recent progress in the metabolic engineering of alkaloids in plant systems
journal, April 2013

  • Glenn, Weslee S.; Runguphan, Weerawat; O’Connor, Sarah E.
  • Current Opinion in Biotechnology, Vol. 24, Issue 2
  • DOI: 10.1016/j.copbio.2012.08.003

Cross-induction of glc and ace Operons of Escherichia coli Attributable to Pathway Intersection : CHARACTERIZATION OF THE
journal, January 1999

  • Pellicer, Maria Teresa; Fernandez, Carmen; Badı́a, Josefa
  • Journal of Biological Chemistry, Vol. 274, Issue 3
  • DOI: 10.1074/jbc.274.3.1745

Direct photosynthetic recycling of carbon dioxide to isobutyraldehyde
journal, November 2009

  • Atsumi, Shota; Higashide, Wendy; Liao, James C.
  • Nature Biotechnology, Vol. 27, Issue 12, p. 1177-1180
  • DOI: 10.1038/nbt.1586

Synthetic non-oxidative glycolysis enables complete carbon conservation
journal, September 2013

  • Bogorad, Igor W.; Lin, Tzu-Shyang; Liao, James C.
  • Nature, Vol. 502, Issue 7473
  • DOI: 10.1038/nature12575

Molecular Characterization of Escherichia coli Malate Synthase G. Differentiation with the Malate Synthase A Isoenzyme
journal, September 1994


Design and analysis of synthetic carbon fixation pathways
journal, April 2010

  • Bar-Even, A.; Noor, E.; Lewis, N. E.
  • Proceedings of the National Academy of Sciences, Vol. 107, Issue 19, p. 8889-8894
  • DOI: 10.1073/pnas.0907176107

Strategies for improving C4 photosynthesis
journal, June 2016


Building carbon–carbon bonds using a biocatalytic methanol condensation cycle
journal, October 2014

  • Bogorad, Igor W.; Chen, Chang-Ting; Theisen, Matthew K.
  • Proceedings of the National Academy of Sciences, Vol. 111, Issue 45
  • DOI: 10.1073/pnas.1413470111

Prospects for improving CO2 fixation in C3-crops through understanding C4-Rubisco biogenesis and catalytic diversity
journal, June 2016

  • Sharwood, Robert E.; Ghannoum, Oula; Whitney, Spencer M.
  • Current Opinion in Plant Biology, Vol. 31
  • DOI: 10.1016/j.pbi.2016.04.002

An Escherichia coli mutant deficient in pyruvate oxidase activity due to altered phospholipid activation of the enzyme
journal, July 1984

  • Chang, Y. -Y.; Cronan, J. E.
  • Proceedings of the National Academy of Sciences, Vol. 81, Issue 14
  • DOI: 10.1073/pnas.81.14.4348

Allosteric control by calcium and mechanism of desensitization of phosphoenolpyruvate carboxykinase of Escherichia coli.
journal, March 1980


Distribution of the Isopropylmalate Pathway to Leucine Among Diverse Bacteria
journal, January 1974


Works referencing / citing this record:

Design and in vitro realization of carbon-conserving photorespiration
journal, November 2018

  • Trudeau, Devin L.; Edlich-Muth, Christian; Zarzycki, Jan
  • Proceedings of the National Academy of Sciences, Vol. 115, Issue 49
  • DOI: 10.1073/pnas.1812605115

Photoreduction of CO 2 with a Formate Dehydrogenase Driven by Photosystem II Using a Semi-artificial Z-Scheme Architecture
journal, November 2018

  • Sokol, Katarzyna P.; Robinson, William E.; Oliveira, Ana R.
  • Journal of the American Chemical Society, Vol. 140, Issue 48
  • DOI: 10.1021/jacs.8b10247

Metabolic rewiring of synthetic pyruvate dehydrogenase bypasses for acetone production in cyanobacteria
journal, February 2020

  • Lee, Hyun Jeong; Son, Jigyeong; Sim, Sang Jun
  • Plant Biotechnology Journal, Vol. 18, Issue 9
  • DOI: 10.1111/pbi.13342

Design and in vitro realization of carbon-conserving photorespiration
journal, November 2018

  • Trudeau, Devin L.; Edlich-Muth, Christian; Zarzycki, Jan
  • Proceedings of the National Academy of Sciences, Vol. 115, Issue 49
  • DOI: 10.1073/pnas.1812605115

Design and in vitro realization of carbon-conserving photorespiration
text, January 2018

  • Trudeau, Devin L.; Edlich-Muth, Christian; Zarzycki, Jan
  • Deutsches Elektronen-Synchrotron, DESY, Hamburg
  • DOI: 10.3204/pubdb-2019-00235

Synthetic biology approaches for improving photosynthesis
journal, January 2019

  • Kubis, Armin; Bar-Even, Arren
  • Journal of Experimental Botany, Vol. 70, Issue 5
  • DOI: 10.1093/jxb/erz029

Figures/Tables have been extracted from DOE-funded journal article accepted manuscripts.