Increased ethylene production by overexpressing phosphoenolpyruvate carboxylase in the cyanobacterium Synechocystis PCC 6803
Abstract
Cyanobacteria can be metabolically engineered to convert CO2 to fuels and chemicals such as ethylene. A major challenge in such efforts is to optimize carbon fixation and partition towards target molecules. The efe gene encoding an ethylene-forming enzyme was introduced into a strain of the cyanobacterium Synechocystis PCC 6803 with increased phosphoenolpyruvate carboxylase (PEPc) levels. The resulting engineered strain (CD-P) showed significantly increased ethylene production (10.5 ± 3.1 ug mL-1 OD-1 day-1) compared to the control strain (6.4 ± 1.4 ug mL-1 OD-1 day-1). Interestingly, extra copies of the native pepc or the heterologous expression of PEPc from the cyanobacterium Synechococcus PCC 7002 (Synechococcus) in the CD-P, increased ethylene production (19.2 ± 1.3 and 18.3 ± 3.3 ug mL-1 OD-1 day-1, respectively) when the cells were treated with the acetyl-CoA carboxylase inhibitor, cycloxydim. A heterologous expression of phosphoenolpyruvate synthase (PPSA) from Synechococcus in the CD-P also increased ethylene production (16.77 ± 4.48 ug mL-1 OD-1 day-1) showing differences in the regulation of the native and the PPSA from Synechococcus in Synechocystis. This work demonstrates that genetic rewiring of cyanobacterial central carbon metabolism can enhance carbon supply to the TCA cycle and thereby further increase ethylene production.
- Authors:
-
- Uppsala Univ. (Sweden)
- National Renewable Energy Laboratory (NREL), Golden, CO (United States)
- Publication Date:
- Research Org.:
- National Renewable Energy Lab. (NREL), Golden, CO (United States)
- Sponsoring Org.:
- USDOE Office of Energy Efficiency and Renewable Energy (EERE), Sustainable Transportation Office. Bioenergy Technologies Office
- OSTI Identifier:
- 1598123
- Report Number(s):
- NREL/JA-2700-74702
Journal ID: ISSN 1754-6834
- Grant/Contract Number:
- AC36-08GO28308
- Resource Type:
- Accepted Manuscript
- Journal Name:
- Biotechnology for Biofuels
- Additional Journal Information:
- Journal Volume: 13; Journal Issue: 1; Journal ID: ISSN 1754-6834
- Publisher:
- BioMed Central
- Country of Publication:
- United States
- Language:
- English
- Subject:
- 09 BIOMASS FUELS; ethylene; phosphoenolpyruvate carboxylase; cyanobacterium; Synechocystis; ALPL
Citation Formats
Durall, Claudia, Lindberg, Pia, Yu, Jianping, and Lindbald, Peter. Increased ethylene production by overexpressing phosphoenolpyruvate carboxylase in the cyanobacterium Synechocystis PCC 6803. United States: N. p., 2020.
Web. doi:10.1186/s13068-020-1653-y.
Durall, Claudia, Lindberg, Pia, Yu, Jianping, & Lindbald, Peter. Increased ethylene production by overexpressing phosphoenolpyruvate carboxylase in the cyanobacterium Synechocystis PCC 6803. United States. https://doi.org/10.1186/s13068-020-1653-y
Durall, Claudia, Lindberg, Pia, Yu, Jianping, and Lindbald, Peter. Tue .
"Increased ethylene production by overexpressing phosphoenolpyruvate carboxylase in the cyanobacterium Synechocystis PCC 6803". United States. https://doi.org/10.1186/s13068-020-1653-y. https://www.osti.gov/servlets/purl/1598123.
@article{osti_1598123,
title = {Increased ethylene production by overexpressing phosphoenolpyruvate carboxylase in the cyanobacterium Synechocystis PCC 6803},
author = {Durall, Claudia and Lindberg, Pia and Yu, Jianping and Lindbald, Peter},
abstractNote = {Cyanobacteria can be metabolically engineered to convert CO2 to fuels and chemicals such as ethylene. A major challenge in such efforts is to optimize carbon fixation and partition towards target molecules. The efe gene encoding an ethylene-forming enzyme was introduced into a strain of the cyanobacterium Synechocystis PCC 6803 with increased phosphoenolpyruvate carboxylase (PEPc) levels. The resulting engineered strain (CD-P) showed significantly increased ethylene production (10.5 ± 3.1 ug mL-1 OD-1 day-1) compared to the control strain (6.4 ± 1.4 ug mL-1 OD-1 day-1). Interestingly, extra copies of the native pepc or the heterologous expression of PEPc from the cyanobacterium Synechococcus PCC 7002 (Synechococcus) in the CD-P, increased ethylene production (19.2 ± 1.3 and 18.3 ± 3.3 ug mL-1 OD-1 day-1, respectively) when the cells were treated with the acetyl-CoA carboxylase inhibitor, cycloxydim. A heterologous expression of phosphoenolpyruvate synthase (PPSA) from Synechococcus in the CD-P also increased ethylene production (16.77 ± 4.48 ug mL-1 OD-1 day-1) showing differences in the regulation of the native and the PPSA from Synechococcus in Synechocystis. This work demonstrates that genetic rewiring of cyanobacterial central carbon metabolism can enhance carbon supply to the TCA cycle and thereby further increase ethylene production.},
doi = {10.1186/s13068-020-1653-y},
journal = {Biotechnology for Biofuels},
number = 1,
volume = 13,
place = {United States},
year = {2020},
month = {1}
}
Web of Science
Works referenced in this record:
Allosteric Inhibition of Phosphoenolpyruvate Carboxylases is Determined by a Single Amino Acid Residue in Cyanobacteria
journal, January 2017
- Takeya, Masahiro; Hirai, Masami Yokota; Osanai, Takashi
- Scientific Reports, Vol. 7, Issue 1
Phototrophic Prokaryotes: The Cyanobacteria
journal, October 1977
- Stanier, R. Y.; Bazine, G. C.
- Annual Review of Microbiology, Vol. 31, Issue 1
Metabolic Engineering of Synechocystis sp. PCC 6803 for Production of the Plant Diterpenoid Manoyl Oxide
journal, July 2015
- Englund, Elias; Andersen-Ranberg, Johan; Miao, Rui
- ACS Synthetic Biology, Vol. 4, Issue 12
Photosynthetic conversion of carbon dioxide to ethylene by the recombinant cyanobacterium, Synechococcus sp. PCC 7942, which harbors a gene for the ethylene-forming enzyme of Pseudomonas syringae
journal, January 1997
- Sakai, Miho; Ogawa, Takahira; Matsuoka, Masayoshi
- Journal of Fermentation and Bioengineering, Vol. 84, Issue 5, p. 434-443
Rubisco: still the most abundant protein of Earth?
journal, February 2013
- Raven, John A.
- New Phytologist, Vol. 198, Issue 1
Bioengineering of carbon fixation, biofuels, and biochemicals in cyanobacteria and plants
journal, November 2012
- Rosgaard, Lisa; de Porcellinis, Alice Jara; Jacobsen, Jacob H.
- Journal of Biotechnology, Vol. 162, Issue 1
Origin and evolution of circadian clock genes in prokaryotes
journal, February 2003
- Dvornyk, V.; Vinogradova, O.; Nevo, E.
- Proceedings of the National Academy of Sciences, Vol. 100, Issue 5
CRISPR-Cas9 for the genome engineering of cyanobacteria and succinate production
journal, November 2016
- Li, Hung; Shen, Claire R.; Huang, Chun-Hung
- Metabolic Engineering, Vol. 38
Sustained photosynthetic conversion of CO2 to ethylene in recombinant cyanobacterium Synechocystis 6803
journal, January 2012
- Ungerer, Justin; Tao, Ling; Davis, Mark
- Energy & Environmental Science, Vol. 5, Issue 10, p. 8998-9006
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
Design, Engineering, and Construction of Photosynthetic Microbial Cell Factories for Renewable Solar Fuel Production
journal, March 2012
- Lindblad, Peter; Lindberg, Pia; Oliveira, Paulo
- AMBIO, Vol. 41, Issue S2
Enhanced stable production of ethylene in photosynthetic cyanobacterium Synechococcus elongatus PCC 7942
journal, May 2019
- Carbonell, Veronica; Vuorio, Eerika; Aro, Eva-Mari
- World Journal of Microbiology and Biotechnology, Vol. 35, Issue 5
Notes: Inhibition of the Acetyl-CoA Carboxylase of Barley Chloroplasts by Cycloxydim and Sethoxydim
journal, December 1987
- Focke, Manfred; Lichtenthaler, Hartmut K.
- Zeitschrift für Naturforschung C, Vol. 42, Issue 11-12
Evaluation of promoters and ribosome binding sites for biotechnological applications in the unicellular cyanobacterium Synechocystis sp. PCC 6803
journal, November 2016
- Englund, Elias; Liang, Feiyan; Lindberg, Pia
- Scientific Reports, Vol. 6, Issue 1
Metabolic engineering of cyanobacteria for the photosynthetic production of succinate
journal, November 2016
- Lan, Ethan I.; Wei, Crystal T.
- Metabolic Engineering, Vol. 38
Development of suitable photobioreactors for CO2 sequestration addressing global warming using green algae and cyanobacteria
journal, April 2011
- Kumar, Kanhaiya; Dasgupta, Chitralekha Nag; Nayak, Bikram
- Bioresource Technology, Vol. 102, Issue 8, p. 4945-4953
Global metabolic rewiring for improved CO2 fixation and chemical production in cyanobacteria
journal, March 2017
- Kanno, Masahiro; Carroll, Austin L.; Atsumi, Shota
- Nature Communications, Vol. 8, Issue 1
Enhancing photosynthetic production of ethylene in genetically engineered Synechocystis sp. PCC 6803
journal, January 2015
- Zhu, Tao; Xie, Xiaoman; Li, Zhimin
- Green Chemistry, Vol. 17, Issue 1
Regulation of glutamine synthetase activity in the unicellular cyanobacterium Synechocystis sp. strain PCC 6803 by the nitrogen source: effect of ammonium.
journal, July 1991
- Mérida, A.; Candau, P.; Florencio, F. J.
- Journal of Bacteriology, Vol. 173, Issue 13
Introduction of a Synthetic CO 2 -fixing Photorespiratory Bypass into a Cyanobacterium
journal, February 2014
- Shih, Patrick M.; Zarzycki, Jan; Niyogi, Krishna K.
- Journal of Biological Chemistry, Vol. 289, Issue 14
Ethylene Biosynthesis
journal, June 1993
- Kende, H.
- Annual Review of Plant Physiology and Plant Molecular Biology, Vol. 44, Issue 1
Ethylene-forming enzyme and bioethylene production
journal, January 2014
- Eckert, Carrie; Xu, Wu; Xiong, Wei
- Biotechnology for Biofuels, Vol. 7, Issue 1
Ethylene Synthesis and Regulated Expression of Recombinant Protein in Synechocystis sp. PCC 6803
journal, November 2012
- Guerrero, Fernando; Carbonell, Verónica; Cossu, Matteo
- PLoS ONE, Vol. 7, Issue 11
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
Metabolome Phenotyping of Inorganic Carbon Limitation in Cells of the Wild Type and Photorespiratory Mutants of the Cyanobacterium Synechocystis sp. Strain PCC 6803
journal, October 2008
- Eisenhut, Marion; Huege, Jan; Schwarz, Doreen
- Plant Physiology, Vol. 148, Issue 4
Mechanisms of carbon fixation and engineering for increased carbon fixation in cyanobacteria
journal, September 2015
- Durall, Claudia; Lindblad, Peter
- Algal Research, Vol. 11
Enhanced growth at low light intensity in the cyanobacterium Synechocystis PCC 6803 by overexpressing phosphoenolpyruvate carboxylase
journal, June 2016
- Durall, Claudia; Rukminasari, Nita; Lindblad, Peter
- Algal Research, Vol. 16
Life and the Evolution of Earth's Atmosphere
journal, May 2002
- Kasting, J. F.
- Science, Vol. 296, Issue 5570
Photosynthetic production of the nitrogen-rich compound guanidine
journal, January 2019
- Wang, Bo; Dong, Tao; Myrlie, Aldon
- Green Chemistry, Vol. 21, Issue 11
Metabolic Changes in Synechocystis PCC6803 upon Nitrogen-Starvation: Excess NADPH Sustains Polyhydroxybutyrate Accumulation
journal, February 2013
- Hauf, Waldemar; Schlebusch, Maximilian; Hüge, Jan
- Metabolites, Vol. 3, Issue 1
Ethylene production with engineered Synechocystis sp PCC 6803 strains
journal, February 2017
- Veetil, Vinod Puthan; Angermayr, S. Andreas; Hellingwerf, Klaas J.
- Microbial Cell Factories, Vol. 16, Issue 1
Construction and analysis of a recombinant cyanobacterium expressing a chromosomally inserted gene for an ethylene-forming enzyme at the psbAI locus
journal, January 2003
- Takahama, Kazutaka; Matsuoka, Masayoshi; Nagahama, Kazuhiro
- Journal of Bioscience and Bioengineering, Vol. 95, Issue 3, p. 302-305
Advances in understanding the cyanobacterial CO2-concentrating-mechanism (CCM): functional components, Ci transporters, diversity, genetic regulation and prospects for engineering into plants
journal, June 2007
- Price, G. Dean; Badger, Murray R.; Woodger, Fiona J.
- Journal of Experimental Botany, Vol. 59, Issue 7
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
Interrelation between Cyanophycin Synthesis, L-Arginine Catabolism and Photosynthesis in the Cyanobacterium Synechocystis Sp. Strain PCC 6803
journal, December 2000
- Stephan, Dirk Paul; Ruppel, Hans Georg; Pistorius, Elfriede K.
- Zeitschrift für Naturforschung C, Vol. 55, Issue 11-12
Engineered cyanobacteria with enhanced growth show increased ethanol production and higher biofuel to biomass ratio
journal, March 2018
- Liang, Feiyan; Englund, Elias; Lindberg, Pia
- Metabolic Engineering, Vol. 46
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
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
Comparison of Ethylene Production by Pseudomonas syringae and Ralstonia solanacearum
journal, May 1999
- Weingart, H.; Völksch, B.; Ullrich, M. S.
- Phytopathology, Vol. 89, Issue 5
Membrane-Inlet Mass Spectrometry Enables a Quantitative Understanding of Inorganic Carbon Uptake Flux and Carbon Concentrating Mechanisms in Metabolically Engineered Cyanobacteria
journal, June 2019
- Douchi, Damien; Liang, Feiyan; Cano, Melissa
- Frontiers in Microbiology, Vol. 10
Enhancement of photosynthetic isobutanol production in engineered cells of Synechocystis PCC 6803
journal, September 2018
- Miao, Rui; Xie, Hao; Lindblad, Peter
- Biotechnology for Biofuels, Vol. 11, Issue 1
Cyanobacteria Perceive Nitrogen Status by Sensing Intracellular 2-Oxoglutarate Levels
journal, October 2001
- Muro-Pastor, M. Isabel; Reyes, José C.; Florencio, Francisco J.
- Journal of Biological Chemistry, Vol. 276, Issue 41
Ethylene biosynthesis
journal, September 1985
- Pirrung, Michael C.
- Bioorganic Chemistry, Vol. 13, Issue 3