Metabolic engineering of Clostridium carboxidivorans for enhanced ethanol and butanol production from syngas and glucose
Abstract
Clostridium carboxidivorans can convert CO2, CO and H2 to ethanol and n-butanol; however, its industrial application is limited by the lack of tools for metabolic pathway engineering. In this study, C. carboxidivorans was successfully engineered to overexpress aor, adhE2, and fnr together with adhE2 or aor. In glucose fermentation, all engineered strains showed higher alcohol yields compared to the wild-type. Strains overexpressing aor showed CO2 re-assimilation during heterotrophic growth. In syngas fermentation, compared to the wild-type, the strain overexpressing adhE2 produced ~50% more ethanol and the strain overexpressing adhE2 and fnr produced ~18% more butanol and ~22% more ethanol. Interestingly, both strains showed obvious acid re-assimilation, with <0.15 g/L total acid remaining at the end of fermentation. Overexpressing fnr with adhE2 enhanced butanol production compared to only adhE2. Furthermore, this is the first report of overexpressing homologous and heterologous genes in C. carboxidivorans for enhancing alcohols production from syngas and glucose.
- Authors:
-
[1];
[2];
[3]
- Dalian Univ. of Technology, Dalian (China); The Ohio State Univ., Columbus, OH (United States)
- The Ohio State Univ., Columbus, OH (United States); Harbin Inst. of Technology, Harbin (China)
- The Ohio State Univ., Columbus, OH (United States)
- Publication Date:
- Research Org.:
- The Ohio State Univ., Columbus, OH (United States)
- Sponsoring Org.:
- USDOE Office of Energy Efficiency and Renewable Energy (EERE)
- OSTI Identifier:
- 1572181
- Alternate Identifier(s):
- OSTI ID: 1548095
- Grant/Contract Number:
- EE0007005
- Resource Type:
- Accepted Manuscript
- Journal Name:
- Bioresource Technology
- Additional Journal Information:
- Journal Volume: 284; Journal Issue: C; Journal ID: ISSN 0960-8524
- Publisher:
- Elsevier
- Country of Publication:
- United States
- Language:
- English
- Subject:
- 09 BIOMASS FUELS; Aldehyde/alcohol dehydrogenase; Aldehyde:ferredoxin oxidoreductase; Clostridium carboxidivorans; Ferredoxin-NAD+ reductase; Syngas fermentation; Metabolic engineering
Citation Formats
Cheng, Chi, Li, Weiming, Lin, Meng, and Yang, Shang-Tian. Metabolic engineering of Clostridium carboxidivorans for enhanced ethanol and butanol production from syngas and glucose. United States: N. p., 2019.
Web. doi:10.1016/j.biortech.2019.03.145.
Cheng, Chi, Li, Weiming, Lin, Meng, & Yang, Shang-Tian. Metabolic engineering of Clostridium carboxidivorans for enhanced ethanol and butanol production from syngas and glucose. United States. https://doi.org/10.1016/j.biortech.2019.03.145
Cheng, Chi, Li, Weiming, Lin, Meng, and Yang, Shang-Tian. Sat .
"Metabolic engineering of Clostridium carboxidivorans for enhanced ethanol and butanol production from syngas and glucose". United States. https://doi.org/10.1016/j.biortech.2019.03.145. https://www.osti.gov/servlets/purl/1572181.
@article{osti_1572181,
title = {Metabolic engineering of Clostridium carboxidivorans for enhanced ethanol and butanol production from syngas and glucose},
author = {Cheng, Chi and Li, Weiming and Lin, Meng and Yang, Shang-Tian},
abstractNote = {Clostridium carboxidivorans can convert CO2, CO and H2 to ethanol and n-butanol; however, its industrial application is limited by the lack of tools for metabolic pathway engineering. In this study, C. carboxidivorans was successfully engineered to overexpress aor, adhE2, and fnr together with adhE2 or aor. In glucose fermentation, all engineered strains showed higher alcohol yields compared to the wild-type. Strains overexpressing aor showed CO2 re-assimilation during heterotrophic growth. In syngas fermentation, compared to the wild-type, the strain overexpressing adhE2 produced ~50% more ethanol and the strain overexpressing adhE2 and fnr produced ~18% more butanol and ~22% more ethanol. Interestingly, both strains showed obvious acid re-assimilation, with <0.15 g/L total acid remaining at the end of fermentation. Overexpressing fnr with adhE2 enhanced butanol production compared to only adhE2. Furthermore, this is the first report of overexpressing homologous and heterologous genes in C. carboxidivorans for enhancing alcohols production from syngas and glucose.},
doi = {10.1016/j.biortech.2019.03.145},
journal = {Bioresource Technology},
number = C,
volume = 284,
place = {United States},
year = {Sat Mar 30 00:00:00 EDT 2019},
month = {Sat Mar 30 00:00:00 EDT 2019}
}
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Works referenced in this record:
Clostridium autoethanogenum, sp. nov., an anaerobic bacterium that produces ethanol from carbon monoxide
journal, April 1994
- Abrini, Jamal; Naveau, Henry; Nyns, Edmond-Jacques
- Archives of Microbiology, Vol. 161, Issue 4
Single gene insertion drives bioalcohol production by a thermophilic archaeon
journal, November 2014
- Basen, Mirko; Schut, Gerrit J.; Nguyen, Diep M.
- Proceedings of the National Academy of Sciences, Vol. 111, Issue 49
CO Metabolism in the Acetogen Acetobacterium woodii
journal, September 2015
- Bertsch, Johannes; Müller, Volker
- Applied and Environmental Microbiology, Vol. 81, Issue 17
Genomic Analysis of Carbon Monoxide Utilization and Butanol Production by Clostridium carboxidivorans Strain P7T
journal, September 2010
- Bruant, Guillaume; Lévesque, Marie-Josée; Peter, Chardeen
- PLoS ONE, Vol. 5, Issue 9
Engineering Clostridium organisms as microbial cell-factories: challenges & opportunities
journal, November 2018
- Charubin, Kamil; Bennett, R. Kyle; Fast, Alan G.
- Metabolic Engineering, Vol. 50
In silico metabolic engineering of Clostridium ljungdahlii for synthesis gas fermentation
journal, November 2016
- Chen, Jin; Henson, Michael A.
- Metabolic Engineering, Vol. 38
Metabolic process engineering of Clostridium tyrobutyricum Δ ack - adhE2 for enhanced n-butanol production from glucose: Effects of methyl viologen on NADH availability, flux distribution, and fermentation kinetics : Metabolic Process Engineering of
journal, December 2014
- Du, Yinming; Jiang, Wenyan; Yu, Mingrui
- Biotechnology and Bioengineering, Vol. 112, Issue 4
Acetogenic mixotrophy: novel options for yield improvement in biofuels and biochemicals production
journal, June 2015
- Fast, Alan G.; Schmidt, Ellinor D.; Jones, Shawn W.
- Current Opinion in Biotechnology, Vol. 33
Efficient butanol-ethanol (B-E) production from carbon monoxide fermentation by Clostridium carboxidivorans
journal, January 2016
- Fernández-Naveira, Ánxela; Abubackar, Haris Nalakath; Veiga, María C.
- Applied Microbiology and Biotechnology, Vol. 100, Issue 7
Glucose bioconversion profile in the syngas-metabolizing species Clostridium carboxidivorans
journal, November 2017
- Fernández-Naveira, Ánxela; Veiga, María C.; Kennes, Christian
- Bioresource Technology, Vol. 244
Production of chemicals from C1 gases (CO, CO2) by Clostridium carboxidivorans
journal, February 2017
- Fernández-Naveira, Ánxela; Abubackar, Haris Nalakath; Veiga, María C.
- World Journal of Microbiology and Biotechnology, Vol. 33, Issue 3
Regulation of Clostridium acetobutylicum metabolism as revealed by mixed-substrate steady-state continuous cultures: role of NADH/NAD ratio and ATP pool.
journal, November 1994
- Girbal, L.; Soucaille, P.
- Journal of Bacteriology, Vol. 176, Issue 21
Acetone production with metabolically engineered strains of Acetobacterium woodii
journal, July 2016
- Hoffmeister, Sabrina; Gerdom, Marzena; Bengelsdorf, Frank R.
- Metabolic Engineering, Vol. 36
Advances in metabolic engineering in the microbial production of fuels and chemicals from C1 gas
journal, April 2018
- Humphreys, Christopher M.; Minton, Nigel P.
- Current Opinion in Biotechnology, Vol. 50
Enhanced Butanol Production Obtained by Reinforcing the Direct Butanol-Forming Route in Clostridium acetobutylicum
journal, October 2012
- Jang, Yu-Sin; Lee, Jin Young; Lee, Joungmin
- mBio, Vol. 3, Issue 5
CO2 fixation by anaerobic non-photosynthetic mixotrophy for improved carbon conversion
journal, September 2016
- Jones, Shawn W.; Fast, Alan G.; Carlson, Ellinor D.
- Nature Communications, Vol. 7, Issue 1
Biological conversion of coal and coal-derived synthesis gas
journal, December 1993
- Klasson, K. T.; Ackerson, M. D.; Clausen, E. C.
- Fuel, Vol. 72, Issue 12, p. 1673-1678
Clostridium ljungdahlii represents a microbial production platform based on syngas
journal, July 2010
- Kopke, M.; Held, C.; Hujer, S.
- Proceedings of the National Academy of Sciences, Vol. 107, Issue 29, p. 13087-13092
Fermentative butanol production by clostridia
journal, October 2008
- Lee, Sang Yup; Park, Jin Hwan; Jang, Seh Hee
- Biotechnology and Bioengineering, Vol. 101, Issue 2, p. 209-228
Biomass, strain engineering, and fermentation processes for butanol production by solventogenic clostridia
journal, August 2016
- Lee, Sang-Hyun; Yun, Eun Ju; Kim, Jungyeon
- Applied Microbiology and Biotechnology, Vol. 100, Issue 19
Complete genome sequence of Clostridium carboxidivorans P7T, a syngas-fermenting bacterium capable of producing long-chain alcohols
journal, October 2015
- Li, Ning; Yang, Junjie; Chai, Changsheng
- Journal of Biotechnology, Vol. 211
Gas Fermentation—A Flexible Platform for Commercial Scale Production of Low-Carbon-Fuels and Chemicals from Waste and Renewable Feedstocks
journal, May 2016
- Liew, FungMin; Martin, Michael E.; Tappel, Ryan C.
- Frontiers in Microbiology, Vol. 7
Metabolic engineering of Clostridium autoethanogenum for selective alcohol production
journal, March 2017
- Liew, Fungmin; Henstra, Anne M.; Kӧpke, Michael
- Metabolic Engineering, Vol. 40
Clostridium carboxidivorans sp. nov., a solvent-producing clostridium isolated from an agricultural settling lagoon, and reclassification of the acetogen Clostridium scatologenes strain SL1 as Clostridium drakei sp. nov.
journal, September 2005
- Liou, J. S. -C.
- INTERNATIONAL JOURNAL OF SYSTEMATIC AND EVOLUTIONARY MICROBIOLOGY, Vol. 55, Issue 5
Low carbon fuels and commodity chemicals from waste gases – systematic approach to understand energy metabolism in a model acetogen
journal, January 2016
- Marcellin, Esteban; Behrendorff, James B.; Nagaraju, Shilpa
- Green Chemistry, Vol. 18, Issue 10
Fixation of CO2 and CO on a diverse range of carbohydrates using anaerobic, non-photosynthetic mixotrophy
journal, February 2018
- Maru, Biniam T.; Munasinghe, Pradeep C.; Gilary, Hadar
- FEMS Microbiology Letters, Vol. 365, Issue 8
Energy Conservation Associated with Ethanol Formation from H 2 and CO 2 in Clostridium autoethanogenum Involving Electron Bifurcation
journal, July 2015
- Mock, Johanna; Zheng, Yanning; Mueller, Alexander P.
- Journal of Bacteriology, Vol. 197, Issue 18
The novel tungsten-iron-sulfur protein of the hyperthermophilic archaebacterium, Pyrococcus furiosus, is an aldehyde ferredoxin oxidoreductase. Evidence for its participation in a unique glycolytic pathway
journal, August 1991
- Mukund, S.; Adams, M. W.
- Journal of Biological Chemistry, Vol. 266, Issue 22
High butanol production by regulating carbon, redox and energy in Clostridia
journal, August 2015
- Ou, Jianfa; Ma, Chao; Xu, Ningning
- Frontiers of Chemical Science and Engineering, Vol. 9, Issue 3
Genome Sequence of the Solvent-Producing Bacterium Clostridium carboxidivorans Strain P7T
journal, August 2010
- Paul, D.; Austin, F. W.; Arick, T.
- Journal of Bacteriology, Vol. 192, Issue 20, p. 5554-5555
Butanol and hexanol production in Clostridium carboxidivorans syngas fermentation: Medium development and culture techniques
journal, August 2015
- Phillips, John R.; Atiyeh, Hasan K.; Tanner, Ralph S.
- Bioresource Technology, Vol. 190
The Complete Genome Sequence of Clostridium aceticum: a Missing Link between Rnf- and Cytochrome-Containing Autotrophic Acetogens
journal, September 2015
- Poehlein, Anja; Cebulla, Martin; Ilg, Marcus M.
- mBio, Vol. 6, Issue 5
Technical guide for genetic advancement of underdeveloped and intractable Clostridium
journal, May 2014
- Pyne, Michael E.; Bruder, Mark; Moo-Young, Murray
- Biotechnology Advances, Vol. 32, Issue 3
Incubation at 25 °C prevents acid crash and enhances alcohol production in Clostridium carboxidivorans P7
journal, September 2015
- Ramió-Pujol, Sara; Ganigué, Ramon; Bañeras, Lluís
- Bioresource Technology, Vol. 192
REBASE—a database for DNA restriction and modification: enzymes, genes and genomes
journal, November 2014
- Roberts, Richard J.; Vincze, Tamas; Posfai, Janos
- Nucleic Acids Research, Vol. 43, Issue D1, p. D298-D299
Autotrophy at the thermodynamic limit of life: a model for energy conservation in acetogenic bacteria
journal, November 2014
- Schuchmann, Kai; Müller, Volker
- Nature Reviews Microbiology, Vol. 12, Issue 12
Syngas fermentation of Clostridium carboxidivoran P7 in a hollow fiber membrane biofilm reactor: Evaluating the mass transfer coefficient and ethanol production performance
journal, April 2014
- Shen, Yanwen; Brown, Robert; Wen, Zhiyou
- Biochemical Engineering Journal, Vol. 85
Enhanced alcohol titre and ratio in carbon monoxide-rich off-gas fermentation of Clostridium carboxidivorans through combination of trace metals optimization with variable-temperature cultivation
journal, September 2017
- Shen, Shaohuang; Gu, Yang; Chai, Changsheng
- Bioresource Technology, Vol. 239
Physiological response of Clostridium carboxidivorans during conversion of synthesis gas to solvents in a gas-fed bioreactor
journal, May 2012
- Ukpong, Michael N.; Atiyeh, Hasan K.; De Lorme, Marthah J. M.
- Biotechnology and Bioengineering, Vol. 109, Issue 11
Engineering clostridia for butanol production from biorenewable resources: from cells to process integration
journal, November 2014
- Wang, Jufang; Yang, Xiaorui; Chen, Chih-Chin
- Current Opinion in Chemical Engineering, Vol. 6
Kinetics of Homoacetic Fermentation of Lactate by Clostridium formicoaceticum
journal, April 1987
- Yang, Shang-Tian; Tang, I-Ching; Okos, Martin R.
- Applied and Environmental Microbiology, Vol. 53, Issue 4
Metabolic and process engineering of Clostridium cellulovorans for biofuel production from cellulose
journal, November 2015
- Yang, Xiaorui; Xu, Mengmeng; Yang, Shang-Tian
- Metabolic Engineering, Vol. 32
Ethanol and acetate production from synthesis gas via fermentation processes using anaerobic bacterium, Clostridium ljungdahlii
journal, December 2005
- Younesi, Habibollah; Najafpour, Ghasem; Mohamed, Abdul Rahman
- Biochemical Engineering Journal, Vol. 27, Issue 2, p. 110-119
Metabolic engineering of Clostridium tyrobutyricum for n-butanol production
journal, July 2011
- Yu, Mingrui; Zhang, Yali; Tang, I-Ching
- Metabolic Engineering, Vol. 13, Issue 4
Effects of different replicons in conjugative plasmids on transformation efficiency, plasmid stability, gene expression and n-butanol biosynthesis in Clostridium tyrobutyricum
journal, December 2011
- Yu, Mingrui; Du, Yinming; Jiang, Wenyan
- Applied Microbiology and Biotechnology, Vol. 93, Issue 2
Metabolic engineering of Clostridium tyrobutyricum for n-butanol production: effects of CoA transferase
journal, April 2015
- Yu, Le; Zhao, Jingbo; Xu, Mengmeng
- Applied Microbiology and Biotechnology, Vol. 99, Issue 11
Effects of end products on fermentation profiles in Clostridium carboxidivorans P7 for syngas fermentation
journal, October 2016
- Zhang, Jie; Taylor, Steven; Wang, Yi
- Bioresource Technology, Vol. 218
Metabolic engineering of Clostridium tyrobutyricum for n-butanol production from sugarcane juice
journal, February 2017
- Zhang, Jianzhi; Yu, Le; Xu, Mengmeng
- Applied Microbiology and Biotechnology, Vol. 101, Issue 10
n-Butanol production from sucrose and sugarcane juice by engineered Clostridium tyrobutyricum overexpressing sucrose catabolism genes and adhE2
journal, June 2017
- Zhang, Jianzhi; Yu, Le; Lin, Meng
- Bioresource Technology, Vol. 233
Works referencing / citing this record:
Experimental Design to Improve Cell Growth and Ethanol Production in Syngas Fermentation by Clostridium carboxidivorans
journal, January 2020
- Benevenuti, Carolina; Botelho, Alanna; Ribeiro, Roberta
- Catalysts, Vol. 10, Issue 1
Metabolic engineering of microorganisms for the production of ethanol and butanol from oxides of carbon
journal, August 2019
- Woo, Ji Eun; Jang, Yu-Sin
- Applied Microbiology and Biotechnology, Vol. 103, Issue 20
Engineering Clostridium for improved solvent production: recent progress and perspective
journal, May 2019
- Cheng, Chi; Bao, Teng; Yang, Shang-Tian
- Applied Microbiology and Biotechnology, Vol. 103, Issue 14
Pathway dissection, regulation, engineering and application: lessons learned from biobutanol production by solventogenic clostridia
journal, March 2020
- Li, Shubo; Huang, Li; Ke, Chengzhu
- Biotechnology for Biofuels, Vol. 13, Issue 1