n-Butanol and ethanol production from cellulose by Clostridium cellulovorans overexpressing heterologous aldehyde/alcohol dehydrogenases

Bao, Teng; Zhao, Jingbo; Li, Jing; Liu, Xin; Yang, Shang-Tian

doi:10.1016/j.biortech.2019.121316

Title: n-Butanol and ethanol production from cellulose by Clostridium cellulovorans overexpressing heterologous aldehyde/alcohol dehydrogenases

Abstract

With high cellulolytic and acetic/butyric acids production abilities, Clostridium cellulovorans is promising for use to produce cellulosic n-butanol. Here, we introduced three different aldehyde/alcohol dehydrogenases encoded by bdhB, adhE1, and adhE2 from Clostridium acetobutylicum into C. cellulovorans and studied their effects on ethanol and n-butanol production. Compared to AdhE2, AdhE1 was more specific for n-butanol biosynthesis over ethanol. Co-expressing adhE1 with bdhB produced a comparable amount of butanol but significantly less ethanol, leading to a high butanol/ethanol ratio of 7.0 and 5.6 (g/g) in glucose and cellulose fermentation, respectively. Co-expressing adhE1 or adhE2 with bdhB did not increase butanol production because the activity of BdhB was limited by the NADPH availability in C. cellulovorans. Altogether, the strain overexpressing adhE2 alone produced the most n-butanol (4.0 g/L, yield: 0.22 ± 0.01 g/g). Based on the insights from this study, further metabolic engineering of C. cellulovorans for cellulosic n-butanol production is suggested.

Authors:

Bao, Teng ^[1]; Zhao, Jingbo ^[1]; Li, Jing ^[2]; Liu, Xin ^[3];

^[1]

The Ohio State Univ., Columbus, OH (United States)
The Ohio State Univ., Columbus, OH (United States); Hebei Univ. of Economics & Business, Shijiazhuang (People's Republic of China)
The Ohio State Univ., Columbus, OH (United States); Changchun Univ. of Technology, Changchun (People's Republic of China)

Publication Date:: 2019-04-03

Research Org.:: The Ohio State Univ., Columbus, OH (United States)

Sponsoring Org.:: USDOE Office of Energy Efficiency and Renewable Energy (EERE)

OSTI Identifier:: 1572179

Alternate Identifier(s):: OSTI ID: 1636742

Grant/Contract Number:: EE0007005

Resource Type:: Accepted Manuscript

Journal Name:: Bioresource Technology

Additional Journal Information:: Journal Volume: 285; Journal Issue: C; Journal ID: ISSN 0960-8524

Publisher:: Elsevier

Country of Publication:: United States

Language:: English

Subject:: 09 BIOMASS FUELS; Biofuel; Clostridium cellulovorans; Consolidated bioprocessing; Metabolic engineering; Butyraldehyde dehydrogenase; Butanol dehydrogenase

Citation Formats


                    Bao, Teng, Zhao, Jingbo, Li, Jing, Liu, Xin, and Yang, Shang-Tian. n-Butanol and ethanol production from cellulose by Clostridium cellulovorans overexpressing heterologous aldehyde/alcohol dehydrogenases.  United States: N. p., 2019. 
Web.  doi:10.1016/j.biortech.2019.121316.

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                    Bao, Teng, Zhao, Jingbo, Li, Jing, Liu, Xin, & Yang, Shang-Tian. n-Butanol and ethanol production from cellulose by Clostridium cellulovorans overexpressing heterologous aldehyde/alcohol dehydrogenases.  United States.  https://doi.org/10.1016/j.biortech.2019.121316

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                    Bao, Teng, Zhao, Jingbo, Li, Jing, Liu, Xin, and Yang, Shang-Tian. Wed .  
"n-Butanol and ethanol production from cellulose by Clostridium cellulovorans overexpressing heterologous aldehyde/alcohol dehydrogenases".  United States.  https://doi.org/10.1016/j.biortech.2019.121316.  https://www.osti.gov/servlets/purl/1572179.

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@article{osti_1572179,

  title        = {n-Butanol and ethanol production from cellulose by Clostridium cellulovorans overexpressing heterologous aldehyde/alcohol dehydrogenases},

  author       = {Bao, Teng and Zhao, Jingbo and Li, Jing and Liu, Xin and Yang, Shang-Tian},

  abstractNote = {With high cellulolytic and acetic/butyric acids production abilities, Clostridium cellulovorans is promising for use to produce cellulosic n-butanol. Here, we introduced three different aldehyde/alcohol dehydrogenases encoded by bdhB, adhE1, and adhE2 from Clostridium acetobutylicum into C. cellulovorans and studied their effects on ethanol and n-butanol production. Compared to AdhE2, AdhE1 was more specific for n-butanol biosynthesis over ethanol. Co-expressing adhE1 with bdhB produced a comparable amount of butanol but significantly less ethanol, leading to a high butanol/ethanol ratio of 7.0 and 5.6 (g/g) in glucose and cellulose fermentation, respectively. Co-expressing adhE1 or adhE2 with bdhB did not increase butanol production because the activity of BdhB was limited by the NADPH availability in C. cellulovorans. Altogether, the strain overexpressing adhE2 alone produced the most n-butanol (4.0 g/L, yield: 0.22 ± 0.01 g/g). Based on the insights from this study, further metabolic engineering of C. cellulovorans for cellulosic n-butanol production is suggested.},

  doi          = {10.1016/j.biortech.2019.121316},

  journal      = {Bioresource Technology},

  number       = C,

  volume       = 285,

  place        = {United States},

  year         = {2019},

  month        = {4}

}

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Title: n-Butanol and ethanol production from cellulose by Clostridium cellulovorans overexpressing heterologous aldehyde/alcohol dehydrogenases

Abstract

Citation Formats

The cellulolytic system of Clostridium cellulolyticum journal, September 1997

Elimination of hydrogenase active site assembly blocks H2 production and increases ethanol yield in Clostridium thermocellum journal, January 2015

Elucidating the contributions of multiple aldehyde/alcohol dehydrogenases to butanol and ethanol production in Clostridium acetobutylicum journal, June 2016

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

Molecular Characterization and Transcriptional Analysis of adhE2, the Gene Encoding the NADH-Dependent Aldehyde/Alcohol Dehydrogenase Responsible for Butanol Production in Alcohologenic Cultures of Clostridium acetobutylicum ATCC 824 journal, February 2002

Metabolic engineering of Clostridium cellulolyticum for the production of n-butanol from crystalline cellulose journal, January 2016

A modular system for Clostridium shuttle plasmids journal, July 2009

Metabolic Engineering of Clostridium cellulolyticum for Production of Isobutanol from Cellulose journal, March 2011

Stable high-titer n-butanol production from sucrose and sugarcane juice by Clostridium acetobutylicum JB200 in repeated batch fermentations journal, July 2014

Acetone-butanol fermentation revisited. journal, January 1986

Recent Developments of the Synthetic Biology Toolkit for Clostridium journal, February 2018

An application programming interface for CellNetAnalyzer journal, August 2011

Characterization of a cellulose-binding, cellulase-containing complex in Clostridium thermocellum. journal, January 1983

Genome-scale reconstruction and in silico analysis of the Clostridium acetobutylicum ATCC 824 metabolic network journal, August 2008

Consolidated bioprocessing of cellulose to isobutanol using Clostridium thermocellum journal, September 2015

Enhanced robustness in acetone-butanol-ethanol fermentation with engineered Clostridium beijerinckii overexpressing adhE2 and ctfAB journal, November 2017

One hundred years of clostridial butanol fermentation journal, January 2016

Genome Sequence and Comparative Analysis of the Solvent-Producing Bacterium Clostridium acetobutylicum journal, August 2001

Recent progress in consolidated bioprocessing journal, June 2012

Process engineering of cellulosic n-butanol production from corn-based biomass using Clostridium cellulovorans journal, November 2017

The Complete Genome Sequence of Clostridium aceticum: a Missing Link between Rnf- and Cytochrome-Containing Autotrophic Acetogens journal, September 2015

Reviving the Weizmann process for commercial n-butanol production journal, September 2018

Advances in consolidated bioprocessing systems for bioethanol and butanol production from biomass: a comprehensive review journal, March 2015

Aldehyde-alcohol dehydrogenase and/or thiolase overexpression coupled with CoA transferase downregulation lead to higher alcohol titers and selectivity in Clostridium acetobutylicum fermentations journal, January 2009

Isolation and Characterization of an Anaerobic, Cellulolytic Bacterium, Clostridium cellulovorans sp. nov journal, January 1984

Genome Sequence of the Cellulosome-Producing Mesophilic Organism Clostridium cellulovorans 743B journal, November 2009

The ald Gene, Encoding a Coenzyme A-Acylating Aldehyde Dehydrogenase, Distinguishes Clostridium beijerinckii and Two Other Solvent-Producing Clostridia fromClostridium acetobutylicum journal, November 1999

Molecular characterization of two Clostridium acetobutylicum ATCC 824 butanol dehydrogenase isozyme genes. journal, January 1992

Engineering clostridia for butanol production from biorenewable resources: from cells to process integration journal, November 2014

Engineering Propionibacterium freudenreichii subsp. shermanii for enhanced propionic acid fermentation: Effects of overexpressing propionyl-CoA:Succinate CoA transferase journal, January 2015

Purification and characterization of the NADH-dependent butanol dehydrogenase from Clostridium acetobutylicum (ATCC 824) journal, September 1989

Enhanced solvent production by metabolic engineering of a twin-clostridial consortium journal, January 2017

Engineering Clostridium acetobutylicum with a histidine kinase knockout for enhanced n-butanol tolerance and production journal, December 2014

High-titer n -butanol production by clostridium acetobutylicum JB200 in fed-batch fermentation with intermittent gas stripping journal, June 2012

Recent advances and state-of-the-art strategies in strain and process engineering for biobutanol production by Clostridium acetobutylicum journal, March 2017

Metabolic and process engineering of Clostridium cellulovorans for biofuel production from cellulose journal, November 2015

Restriction modification system analysis and development of in vivo methylation for the transformation of Clostridium cellulovorans journal, November 2015

A Quantitative System-Scale Characterization of the Metabolism of Clostridium acetobutylicum journal, November 2015

Elucidation of the roles of adhE1 and adhE2 in the primary metabolism of Clostridium acetobutylicum by combining in-frame gene deletion and a quantitative system-scale approach journal, April 2016

Metabolic flexibility of a butyrate pathway mutant of Clostridium acetobutylicum journal, March 2017

Metabolic engineering of Clostridium tyrobutyricum for n-butanol production journal, July 2011

Metabolic engineering of Clostridium tyrobutyricum for n-butanol production from maltose and soluble starch by overexpressing α-glucosidase journal, May 2015

Metabolic engineering of Clostridium tyrobutyricum for n-butanol production through co-utilization of glucose and xylose : Metabolic Engineering of journal, June 2015

n-Butanol production from sucrose and sugarcane juice by engineered Clostridium tyrobutyricum overexpressing sucrose catabolism genes and adhE2 journal, June 2017

Exploiting endogenous CRISPR-Cas system for multiplex genome editing in Clostridium tyrobutyricum and engineer the strain for high-level butanol production journal, May 2018

Metabolic engineering strategies for consolidated production of lactic acid from lignocellulosic biomass journal, January 2020

Metabolic and evolutionary responses of Clostridium thermocellum to genetic interventions aimed at improving ethanol production journal, March 2020

Consolidated bioprocessing for butanol production of cellulolytic Clostridia: development and optimization journal, August 2019

Development of a shuttle plasmid without host restriction sites for efficient transformation and heterologous gene expression in Clostridium cellulovorans journal, May 2019

Engineering Clostridium for improved solvent production: recent progress and perspective journal, May 2019

The cellulolytic system of Clostridium cellulolyticum
journal, September 1997

Elimination of hydrogenase active site assembly blocks H2 production and increases ethanol yield in Clostridium thermocellum
journal, January 2015

Elucidating the contributions of multiple aldehyde/alcohol dehydrogenases to butanol and ethanol production in Clostridium acetobutylicum
journal, June 2016

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

Molecular Characterization and Transcriptional Analysis of adhE2, the Gene Encoding the NADH-Dependent Aldehyde/Alcohol Dehydrogenase Responsible for Butanol Production in Alcohologenic Cultures of Clostridium acetobutylicum ATCC 824
journal, February 2002

Metabolic engineering of Clostridium cellulolyticum for the production of n-butanol from crystalline cellulose
journal, January 2016

A modular system for Clostridium shuttle plasmids
journal, July 2009

Metabolic Engineering of Clostridium cellulolyticum for Production of Isobutanol from Cellulose
journal, March 2011

Stable high-titer n-butanol production from sucrose and sugarcane juice by Clostridium acetobutylicum JB200 in repeated batch fermentations
journal, July 2014

Acetone-butanol fermentation revisited.
journal, January 1986

Recent Developments of the Synthetic Biology Toolkit for Clostridium
journal, February 2018

An application programming interface for CellNetAnalyzer
journal, August 2011

Characterization of a cellulose-binding, cellulase-containing complex in Clostridium thermocellum.
journal, January 1983

Genome-scale reconstruction and in silico analysis of the Clostridium acetobutylicum ATCC 824 metabolic network
journal, August 2008

Consolidated bioprocessing of cellulose to isobutanol using Clostridium thermocellum
journal, September 2015

Enhanced robustness in acetone-butanol-ethanol fermentation with engineered Clostridium beijerinckii overexpressing adhE2 and ctfAB
journal, November 2017

One hundred years of clostridial butanol fermentation
journal, January 2016

Genome Sequence and Comparative Analysis of the Solvent-Producing Bacterium Clostridium acetobutylicum
journal, August 2001

Recent progress in consolidated bioprocessing
journal, June 2012

Process engineering of cellulosic n-butanol production from corn-based biomass using Clostridium cellulovorans
journal, November 2017

The Complete Genome Sequence of Clostridium aceticum: a Missing Link between Rnf- and Cytochrome-Containing Autotrophic Acetogens
journal, September 2015

Reviving the Weizmann process for commercial n-butanol production
journal, September 2018

Advances in consolidated bioprocessing systems for bioethanol and butanol production from biomass: a comprehensive review
journal, March 2015

Aldehyde-alcohol dehydrogenase and/or thiolase overexpression coupled with CoA transferase downregulation lead to higher alcohol titers and selectivity in Clostridium acetobutylicum fermentations
journal, January 2009

Isolation and Characterization of an Anaerobic, Cellulolytic Bacterium, Clostridium cellulovorans sp. nov
journal, January 1984

Genome Sequence of the Cellulosome-Producing Mesophilic Organism Clostridium cellulovorans 743B
journal, November 2009

The ald Gene, Encoding a Coenzyme A-Acylating Aldehyde Dehydrogenase, Distinguishes Clostridium beijerinckii and Two Other Solvent-Producing Clostridia fromClostridium acetobutylicum
journal, November 1999

Molecular characterization of two Clostridium acetobutylicum ATCC 824 butanol dehydrogenase isozyme genes.
journal, January 1992

Engineering clostridia for butanol production from biorenewable resources: from cells to process integration
journal, November 2014

Engineering Propionibacterium freudenreichii subsp. shermanii for enhanced propionic acid fermentation: Effects of overexpressing propionyl-CoA:Succinate CoA transferase
journal, January 2015

Purification and characterization of the NADH-dependent butanol dehydrogenase from Clostridium acetobutylicum (ATCC 824)
journal, September 1989

Enhanced solvent production by metabolic engineering of a twin-clostridial consortium
journal, January 2017

Engineering Clostridium acetobutylicum with a histidine kinase knockout for enhanced n-butanol tolerance and production
journal, December 2014

High-titer n -butanol production by clostridium acetobutylicum JB200 in fed-batch fermentation with intermittent gas stripping
journal, June 2012

Recent advances and state-of-the-art strategies in strain and process engineering for biobutanol production by Clostridium acetobutylicum
journal, March 2017

Metabolic and process engineering of Clostridium cellulovorans for biofuel production from cellulose
journal, November 2015

Restriction modification system analysis and development of in vivo methylation for the transformation of Clostridium cellulovorans
journal, November 2015

A Quantitative System-Scale Characterization of the Metabolism of Clostridium acetobutylicum
journal, November 2015

Elucidation of the roles of adhE1 and adhE2 in the primary metabolism of Clostridium acetobutylicum by combining in-frame gene deletion and a quantitative system-scale approach
journal, April 2016

Metabolic flexibility of a butyrate pathway mutant of Clostridium acetobutylicum
journal, March 2017

Metabolic engineering of Clostridium tyrobutyricum for n-butanol production
journal, July 2011

Metabolic engineering of Clostridium tyrobutyricum for n-butanol production from maltose and soluble starch by overexpressing α-glucosidase
journal, May 2015

Metabolic engineering of Clostridium tyrobutyricum for n-butanol production through co-utilization of glucose and xylose : Metabolic Engineering of
journal, June 2015

n-Butanol production from sucrose and sugarcane juice by engineered Clostridium tyrobutyricum overexpressing sucrose catabolism genes and adhE2
journal, June 2017

Exploiting endogenous CRISPR-Cas system for multiplex genome editing in Clostridium tyrobutyricum and engineer the strain for high-level butanol production
journal, May 2018

Metabolic engineering strategies for consolidated production of lactic acid from lignocellulosic biomass
journal, January 2020

Metabolic and evolutionary responses of Clostridium thermocellum to genetic interventions aimed at improving ethanol production
journal, March 2020

Consolidated bioprocessing for butanol production of cellulolytic Clostridia: development and optimization
journal, August 2019

Development of a shuttle plasmid without host restriction sites for efficient transformation and heterologous gene expression in Clostridium cellulovorans
journal, May 2019

Engineering Clostridium for improved solvent production: recent progress and perspective
journal, May 2019