Cyanobacterial carbon metabolism: Fluxome plasticity and oxygen dependence: Cyanobacterial Carbon Metabolism
Abstract
ABSTRACT Synechocystis sp. strain PCC 6803 has been widely used as a photo‐biorefinery chassis. Based on its genome annotation, this species contains a complete TCA cycle, an Embden‐Meyerhof‐Parnas pathway (EMPP), an oxidative pentose phosphate pathway (OPPP), and an Entner–Doudoroff pathway (EDP). To evaluate how Synechocystis 6803 catabolizes glucose under heterotrophic conditions, we performed 13 C metabolic flux analysis, metabolite pool size analysis, gene knockouts, and heterologous expressions. The results revealed a cyclic mode of flux through the OPPP. Small, but non‐zero, fluxes were observed through the TCA cycle and the malic shunt. Independent knockouts of 6‐phosphogluconate dehydrogenase ( gnd ) and malic enzyme ( me ) corroborated these results, as neither mutant could grow under dark heterotrophic conditions. Our data also indicate that Synechocystis 6803 metabolism relies upon oxidative phosphorylation to generate ATP from NADPH under dark or insufficient light conditions. The pool sizes of intermediates in the TCA cycle, particularly acetyl‐CoA, were found to be several fold lower in Synechocystis 6803 (compared to E. coli metabolite pool sizes), while its sugar phosphate intermediates were several‐fold higher. Moreover, negligible flux was detected through the native, or heterologous, EDP in the wild type or Δ gnd strains under heterotrophic conditions. Comparingmore »
- Authors:
-
- Washington Univ., St. Louis, MO (United States)
- Joint BioEnergy Inst. (JBEI), Emeryville, CA (United States); Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States)
- Joint BioEnergy Inst. (JBEI), Emeryville, CA (United States); Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States); Univ. of California, Berkeley, CA (United States); Technical Univ. of Denmark, Lyngby (Denmark)
- Department of Energy, Environmental and Chemical Engineering, Washington University in St. Louis, St. Louis Missouri 63130
- Publication Date:
- Research Org.:
- Lawrence Berkeley National Laboratory (LBNL), Berkeley, CA (United States)
- Sponsoring Org.:
- USDOE Office of Science (SC), Biological and Environmental Research (BER)
- OSTI Identifier:
- 1393244
- Alternate Identifier(s):
- OSTI ID: 1401285
- Grant/Contract Number:
- AC02-05CH11231; SCGF2015; DESC0012722
- Resource Type:
- Accepted Manuscript
- Journal Name:
- Biotechnology and Bioengineering
- Additional Journal Information:
- Journal Volume: 114; Journal Issue: 7; Journal ID: ISSN 0006-3592
- Publisher:
- Wiley
- Country of Publication:
- United States
- Language:
- English
- Subject:
- 59 BASIC BIOLOGICAL SCIENCES
Citation Formats
Wan, Ni, DeLorenzo, Drew M., He, Lian, You, Le, Immethun, Cheryl M., Wang, George, Baidoo, Edward E. K., Hollinshead, Whitney, Keasling, Jay D., Moon, Tae Seok, and Tang, Yinjie J. Cyanobacterial carbon metabolism: Fluxome plasticity and oxygen dependence: Cyanobacterial Carbon Metabolism. United States: N. p., 2017.
Web. doi:10.1002/bit.26287.
Wan, Ni, DeLorenzo, Drew M., He, Lian, You, Le, Immethun, Cheryl M., Wang, George, Baidoo, Edward E. K., Hollinshead, Whitney, Keasling, Jay D., Moon, Tae Seok, & Tang, Yinjie J. Cyanobacterial carbon metabolism: Fluxome plasticity and oxygen dependence: Cyanobacterial Carbon Metabolism. United States. https://doi.org/10.1002/bit.26287
Wan, Ni, DeLorenzo, Drew M., He, Lian, You, Le, Immethun, Cheryl M., Wang, George, Baidoo, Edward E. K., Hollinshead, Whitney, Keasling, Jay D., Moon, Tae Seok, and Tang, Yinjie J. Thu .
"Cyanobacterial carbon metabolism: Fluxome plasticity and oxygen dependence: Cyanobacterial Carbon Metabolism". United States. https://doi.org/10.1002/bit.26287. https://www.osti.gov/servlets/purl/1393244.
@article{osti_1393244,
title = {Cyanobacterial carbon metabolism: Fluxome plasticity and oxygen dependence: Cyanobacterial Carbon Metabolism},
author = {Wan, Ni and DeLorenzo, Drew M. and He, Lian and You, Le and Immethun, Cheryl M. and Wang, George and Baidoo, Edward E. K. and Hollinshead, Whitney and Keasling, Jay D. and Moon, Tae Seok and Tang, Yinjie J.},
abstractNote = {ABSTRACT Synechocystis sp. strain PCC 6803 has been widely used as a photo‐biorefinery chassis. Based on its genome annotation, this species contains a complete TCA cycle, an Embden‐Meyerhof‐Parnas pathway (EMPP), an oxidative pentose phosphate pathway (OPPP), and an Entner–Doudoroff pathway (EDP). To evaluate how Synechocystis 6803 catabolizes glucose under heterotrophic conditions, we performed 13 C metabolic flux analysis, metabolite pool size analysis, gene knockouts, and heterologous expressions. The results revealed a cyclic mode of flux through the OPPP. Small, but non‐zero, fluxes were observed through the TCA cycle and the malic shunt. Independent knockouts of 6‐phosphogluconate dehydrogenase ( gnd ) and malic enzyme ( me ) corroborated these results, as neither mutant could grow under dark heterotrophic conditions. Our data also indicate that Synechocystis 6803 metabolism relies upon oxidative phosphorylation to generate ATP from NADPH under dark or insufficient light conditions. The pool sizes of intermediates in the TCA cycle, particularly acetyl‐CoA, were found to be several fold lower in Synechocystis 6803 (compared to E. coli metabolite pool sizes), while its sugar phosphate intermediates were several‐fold higher. Moreover, negligible flux was detected through the native, or heterologous, EDP in the wild type or Δ gnd strains under heterotrophic conditions. Comparing photoautotrophic, photomixotrophic, and heterotrophic conditions, the Calvin cycle, OPPP, and EMPP in Synechocystis 6803 possess the ability to regulate their fluxes under various growth conditions (plastic), whereas its TCA cycle always maintains at low levels (rigid). This work also demonstrates how genetic profiles do not always reflect actual metabolic flux through native or heterologous pathways. Biotechnol. Bioeng. 2017;114: 1593–1602. © 2017 Wiley Periodicals, Inc.},
doi = {10.1002/bit.26287},
journal = {Biotechnology and Bioengineering},
number = 7,
volume = 114,
place = {United States},
year = {Thu Mar 30 00:00:00 EDT 2017},
month = {Thu Mar 30 00:00:00 EDT 2017}
}
Web of Science
Works referenced in this record:
The Malic Enzyme Is Required for Optimal Photoautotrophic Growth of Synechocystis sp. Strain PCC 6803 under Continuous Light but Not under a Diurnal Light Regimen
journal, November 2004
- Bricker, T. M.; Zhang, S.; Laborde, S. M.
- Journal of Bacteriology, Vol. 186, Issue 23
The Calvin cycle in cyanobacteria is regulated by CP12 via the NAD(H)/NADP(H) ratio under light/dark conditions: Regulation of the Calvin cycle by cyanobacterial CP12
journal, April 2005
- Tamoi, Masahiro; Miyazaki, Takashi; Fukamizo, Tamo
- The Plant Journal, Vol. 42, Issue 4
Engineering nonphosphorylative metabolism to generate lignocellulose-derived products
journal, February 2016
- Tai, Yi-Shu; Xiong, Mingyong; Jambunathan, Pooja
- Nature Chemical Biology, Vol. 12, Issue 4
Highly expressed and alien genes of the Synechocystis genome
journal, April 2001
- Mrazek, J.
- Nucleic Acids Research, Vol. 29, Issue 7
Not just a circle: flux modes in the plant TCA cycle
journal, August 2010
- Sweetlove, Lee J.; Beard, Katherine F. M.; Nunes-Nesi, Adriano
- Trends in Plant Science, Vol. 15, Issue 8
Metabolic engineering of cyanobacteria for the synthesis of commodity products
journal, June 2015
- Angermayr, S. Andreas; Gorchs Rovira, Aleix; Hellingwerf, Klaas J.
- Trends in Biotechnology, Vol. 33, Issue 6
Development of Synechocystis sp. PCC 6803 as a Phototrophic Cell Factory
journal, August 2013
- Yu, Yi; You, Le; Liu, Dianyi
- Marine Drugs, Vol. 11, Issue 8
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
Reconstruction and analysis of genome-scale metabolic model of a photosynthetic bacterium
journal, November 2010
- Montagud, Arnau; Navarro, Emilio; Fernández de Córdoba, Pedro
- BMC Systems Biology, Vol. 4, Issue 1
Synthetic biology of cyanobacteria: unique challenges and opportunities
journal, January 2013
- Berla, Bertram M.; Saha, Rajib; Immethun, Cheryl M.
- Frontiers in Microbiology, Vol. 4
The γ-aminobutyric acid shunt contributes to closing the tricarboxylic acid cycle in S ynechocystis sp. PCC 6803 : The γ-aminobutyric acid shunt in
journal, July 2014
- Xiong, Wei; Brune, Daniel; Vermaas, Wim F. J.
- Molecular Microbiology, Vol. 93, Issue 4
Flux Balance Analysis of Photoautotrophic Metabolism
journal, December 2005
- Shastri, A. A.; Morgan, J. A.
- Biotechnology Progress, Vol. 21, Issue 6
Cyanobacterial photo-driven mixotrophic metabolism and its advantages for biosynthesis
journal, July 2015
- Wan, Ni; Abernathy, Mary; Tang, Joseph Kuo-Hsiang
- Frontiers of Chemical Science and Engineering, Vol. 9, Issue 3
Absolute quantitation of intracellular metabolite concentrations by an isotope ratio-based approach
journal, July 2008
- Bennett, Bryson D.; Yuan, Jie; Kimball, Elizabeth H.
- Nature Protocols, Vol. 3, Issue 8
Rapid metabolic analysis of Rhodococcus opacus PD630 via parallel 13 C-metabolite fingerprinting
journal, September 2015
- Hollinshead, Whitney D.; Henson, William R.; Abernathy, Mary
- Biotechnology and Bioengineering, Vol. 113, Issue 1
Photoheterotrophic Fluxome in Synechocystis sp. Strain PCC 6803 and Its Implications for Cyanobacterial Bioenergetics
journal, December 2014
- You, Le; He, Lian; Tang, Yinjie J.
- Journal of Bacteriology, Vol. 197, Issue 5
Metabolic flux analysis with a comprehensive isotopomer model inBacillus subtilis
journal, January 2001
- Dauner, Michael; Bailey, James E.; Sauer, Uwe
- Biotechnology and Bioengineering, Vol. 76, Issue 2
Integrated Metabolic Flux and Omics Analysis of Synechocystis sp. PCC 6803 under Mixotrophic and Photoheterotrophic Conditions
journal, June 2014
- Nakajima, Tsubasa; Kajihata, Shuichi; Yoshikawa, Katsunori
- Plant and Cell Physiology, Vol. 55, Issue 9
Diurnal Regulation of Cellular Processes in the Cyanobacterium Synechocystis sp. Strain PCC 6803: Insights from Transcriptomic, Fluxomic, and Physiological Analyses
journal, May 2016
- Saha, Rajib; Liu, Deng; Hoynes-O’Connor, Allison
- mBio, Vol. 7, Issue 3
Oxygen-responsive genetic circuits constructed in Synechocystis sp. PCC 6803 : Oxygen-Responsive Gene Circuits for
journal, September 2015
- Immethun, Cheryl M.; Ng, Kenneth M.; DeLorenzo, Drew M.
- Biotechnology and Bioengineering, Vol. 113, Issue 2
Overflow Metabolism in Escherichia coli during Steady-State Growth: Transcriptional Regulation and Effect of the Redox Ratio
journal, May 2006
- Vemuri, G. N.; Altman, E.; Sangurdekar, D. P.
- Applied and Environmental Microbiology, Vol. 72, Issue 5
Metabolic Flux Analysis in Synechocystis Using Isotope Distribution from 13C-Labeled Glucose
journal, July 2002
- Yang, C.
- Metabolic Engineering, Vol. 4, Issue 3
13 C-MFA delineates the photomixotrophic metabolism of Synechocystis sp. PCC 6803 under light- and carbon-sufficient conditions
journal, April 2014
- You, Le; Berla, Bert; He, Lian
- Biotechnology Journal, Vol. 9, Issue 5
Design and analysis of metabolic pathways supporting formatotrophic growth for electricity-dependent cultivation of microbes
journal, August 2013
- Bar-Even, Arren; Noor, Elad; Flamholz, Avi
- Biochimica et Biophysica Acta (BBA) - Bioenergetics, Vol. 1827, Issue 8-9
The plasticity of cyanobacterial metabolism supports direct CO2 conversion to ethylene
journal, April 2015
- Xiong, Wei; Morgan, John A.; Ungerer, Justin
- Nature Plants, Vol. 1, Issue 5
Rethinking glycolysis: on the biochemical logic of metabolic pathways
journal, May 2012
- Bar-Even, Arren; Flamholz, Avi; Noor, Elad
- Nature Chemical Biology, Vol. 8, Issue 6
Effect of malic enzyme on ethanol production by Synechocystis sp. PCC 6803
journal, January 2015
- Yoshikawa, Katsunori; Hirasawa, Takashi; Shimizu, Hiroshi
- Journal of Bioscience and Bioengineering, Vol. 119, Issue 1
Phosphoketolase pathway contributes to carbon metabolism in cyanobacteria
journal, December 2015
- Xiong, Wei; Lee, Tai-Chi; Rommelfanger, Sarah
- Nature Plants, Vol. 2, Issue 1, Article No. 15187
Mapping photoautotrophic metabolism with isotopically nonstationary 13C flux analysis
journal, November 2011
- Young, Jamey D.; Shastri, Avantika A.; Stephanopoulos, Gregory
- Metabolic Engineering, Vol. 13, Issue 6
Characterization of a novel unconjugated pteridine glycoside, cyanopterin, in Synechocystis sp. PCC 6803
journal, January 1999
- Lee, Hee Woo; Oh, Chang Ho; Geyer, Armin
- Biochimica et Biophysica Acta (BBA) - Bioenergetics, Vol. 1410, Issue 1
Role of plant glyoxylate reductases during stress: a hypothesis
journal, September 2009
- Allan, Wendy L.; Clark, Shawn M.; Hoover, Gordon J.
- Biochemical Journal, Vol. 423, Issue 1
Light-activated heterotrophic growth of the cyanobacterium Synechocystis sp. strain PCC 6803: a blue-light-requiring process.
journal, May 1991
- Anderson, S. L.; McIntosh, L.
- Journal of Bacteriology, Vol. 173, Issue 9
A systems biology approach to reconcile metabolic network models with application to Synechocystis sp. PCC 6803 for biofuel production
journal, January 2016
- Mohammadi, Reza; Fallah-Mehrabadi, Jalil; Bidkhori, Gholamreza
- Molecular BioSystems, Vol. 12, Issue 8
Detailing the optimality of photosynthesis in cyanobacteria through systems biology analysis
journal, January 2012
- Nogales, J.; Gudmundsson, S.; Knight, E. M.
- Proceedings of the National Academy of Sciences, Vol. 109, Issue 7
Sll1330 controls the expression of glycolytic genes in Synechocystis sp. PCC 6803
journal, April 2007
- Tabei, Yosuke; Okada, Katsuhiko; Tsuzuki, Mikio
- Biochemical and Biophysical Research Communications, Vol. 355, Issue 4
Accurate Assessment of Amino Acid Mass Isotopomer Distributions for Metabolic Flux Analysis
journal, October 2007
- Antoniewicz, Maciek R.; Kelleher, Joanne K.; Stephanopoulos, Gregory
- Analytical Chemistry, Vol. 79, Issue 19
Engineering cyanobacteria to generate high-value products
journal, February 2011
- Ducat, Daniel C.; Way, Jeffrey C.; Silver, Pamela A.
- Trends in Biotechnology, Vol. 29, Issue 2, p. 95-103
The Entner–Doudoroff pathway is an overlooked glycolytic route in cyanobacteria and plants
journal, April 2016
- Chen, Xi; Schreiber, Karoline; Appel, Jens
- Proceedings of the National Academy of Sciences, Vol. 113, Issue 19
New tools for mass isotopomer data evaluation in13C flux analysis: Mass isotope correction, data consistency checking, and precursor relationships
journal, January 2004
- Wahl, S. Aljoscha; Dauner, Michael; Wiechert, Wolfgang
- Biotechnology and Bioengineering, Vol. 85, Issue 3
MIRACLE: mass isotopomer ratio analysis of U-13C-labeled extracts. A new method for accurate quantification of changes in concentrations of intracellular metabolites
journal, January 2004
- Mashego, M. R.; Wu, L.; Van Dam, J. C.
- Biotechnology and Bioengineering, Vol. 85, Issue 6
Enhanced Survival of the Cyanobacterium Oscillatoria terebriformis in Darkness under Anaerobic Conditions †
journal, January 1987
- Richardson, Laurie L.; Castenholz, Richard W.
- Applied and Environmental Microbiology, Vol. 53, Issue 9
Works referencing / citing this record:
Engineering photosynthetic production of L-lysine
journal, November 2017
- Korosh, Travis C.; Markley, Andrew L.; Clark, Ryan L.
- Metabolic Engineering, Vol. 44
Citrate synthase from Synechocystis is a distinct class of bacterial citrate synthase
journal, April 2019
- Ito, Shoki; Koyama, Naoto; Osanai, Takashi
- Scientific Reports, Vol. 9, Issue 1
Biochemical characterisation of fumarase C from a unicellular cyanobacterium demonstrating its substrate affinity, altered by an amino acid substitution
journal, July 2019
- Katayama, Noriaki; Takeya, Masahiro; Osanai, Takashi
- Scientific Reports, Vol. 9, Issue 1
Metabolic pathway rewiring in engineered cyanobacteria for solar-to-chemical and solar-to-fuel production from CO2
journal, May 2017
- Woo, Han Min
- Bioengineered, Vol. 9, Issue 1
13C-Metabolic Flux Analysis Reveals the Metabolic Flux Redistribution for Enhanced Production of Poly-γ-Glutamic Acid in dlt Over-Expressed Bacillus licheniformis
journal, February 2019
- He, Penghui; Wan, Ni; Cai, Dongbo
- Frontiers in Microbiology, Vol. 10
Purification and Characterisation of Malate Dehydrogenase From Synechocystis sp. PCC 6803: Biochemical Barrier of the Oxidative Tricarboxylic Acid Cycle
journal, July 2018
- Takeya, Masahiro; Ito, Shoki; Sukigara, Haruna
- Frontiers in Plant Science, Vol. 9