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

Du, Yinming; Jiang, Wenyan; Yu, Mingrui; Tang, I‐Ching; Yang, Shang‐Tian

doi:10.1002/bit.25489

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

Journal Article · Tue Dec 23 00:00:00 EST 2014 · Biotechnology and Bioengineering

DOI:https://doi.org/10.1002/bit.25489· OSTI ID:1400884

Du, Yinming ^[1]; Jiang, Wenyan ^[1]; Yu, Mingrui ^[2]; Tang, I‐Ching ^[2]; Yang, Shang‐Tian ^[1]

William G. Lowrie Department of Chemical and Biomolecular Engineering The Ohio State University 140 West 19th Avenue Columbus Ohio 43210
Bioprocessing Innovative Company Dublin Ohio

ABSTRACT

Butanol biosynthesis through aldehyde/alcohol dehydrogenase ( adhE2 ) is usually limited by NADH availability, resulting in low butanol titer, yield, and productivity. To alleviate this limitation and improve n‐butanol production by Clostridium tyrobutyricum Δ ack – adhE2 overexpressing adhE2 , the NADH availability was increased by using methyl viologen (MV) as an artificial electron carrier to divert electrons from ferredoxin normally used for H ₂ production. In the batch fermentation with the addition of 500 μM MV, H ₂ , acetate, and butyrate production was reduced by more than 80–90%, while butanol production increased more than 40% to 14.5 g/L. Metabolic flux analysis revealed that butanol production increased in the fermentation with MV because of increased NADH availability as a result of reduced H ₂ production. Furthermore, continuous butanol production of ∼55 g/L with a high yield of ∼0.33 g/g glucose and extremely low ethanol, acetate, and butyrate production was obtained in fed‐batch fermentation with gas stripping for in situ butanol recovery. This study demonstrated a stable and reliable process for high‐yield and high‐titer n‐butanol production by metabolically engineered C. tyrobutyricum by applying MV as an electron carrier to increase butanol biosynthesis. Biotechnol. Bioeng. 2015;112: 705–715. © 2014 Wiley Periodicals, Inc.

View Accepted Manuscript (Publisher)

Sponsoring Organization:: USDOE

Grant/Contract Number:: AR0000095

OSTI ID:: 1400884

Journal Information:: Biotechnology and Bioengineering, Journal Name: Biotechnology and Bioengineering Journal Issue: 4 Vol. 112; ISSN 0006-3592

Publisher:: Wiley Blackwell (John Wiley & Sons)Copyright Statement

Country of Publication:: United States

Language:: English

References (45)

Modulation of metabolism ofClostridium acetobutylicum grown in chemostat culture in a three-electrode potentiostatic system with methyl viologen as electron carrier Peguin, Sophie; Soucaille, Philippe Biotechnology and Bioengineering, Vol. 51, Issue 3 https://doi.org/10.1002/(SICI)1097-0290(19960805)51:3<342::AID-BIT9>3.0.CO;2-D	journal	August 1996
Fermentative butanol production by clostridia Lee, Sang Yup; Park, Jin Hwan; Jang, Seh Hee Biotechnology and Bioengineering, Vol. 101, Issue 2, p. 209-228 https://doi.org/10.1002/bit.22003	journal	October 2008
Butyrate production enhancement by Clostridium tyrobutyricum using electron mediators and a cathodic electron donor Choi, Okkyoung; Um, Youngsoon; Sang, Byoung-In Biotechnology and Bioengineering, Vol. 109, Issue 10 https://doi.org/10.1002/bit.24520	journal	April 2012
High-titer n -butanol production by clostridium acetobutylicum JB200 in fed-batch fermentation with intermittent gas stripping Xue, Chuang; Zhao, Jingbo; Lu, Congcong Biotechnology and Bioengineering, Vol. 109, Issue 11 https://doi.org/10.1002/bit.24563	journal	June 2012
Agitation and pressure effects on acetone-butanol fermentation Doremus, Michael G.; Linden, James C.; Moreira, Antonio R. Biotechnology and Bioengineering, Vol. 27, Issue 6 https://doi.org/10.1002/bit.260270615	journal	June 1985
Importance of agitation in acetone-butanol fermentation Yerushalmi, Laleh; Volesky, Bohumil Biotechnology and Bioengineering, Vol. 27, Issue 9 https://doi.org/10.1002/bit.260270905	journal	September 1985
Metabolic flexibility of Clostridium acetobutylicum in response to methyl viologen addition Peguin, S.; Goma, G.; Delorme, P. Applied Microbiology and Biotechnology, Vol. 42, Issue 4 https://doi.org/10.1007/BF00173928	journal	December 1994
Enzymatic investigations on butanol dehydrogenase and butyraldehyde dehydrogenase in extracts of Clostridium acetobutylicum D�rre, Peter; Kuhn, Anita; Gottwald, Matthias Applied Microbiology and Biotechnology, Vol. 26, Issue 3 https://doi.org/10.1007/BF00286322	journal	June 1987
Electron flow shift inClostridium acetobutylicum fermentation by electrochemically introduced reducing equivalent Kim, Tae Sung; Kim, Byung Hong Biotechnology Letters, Vol. 10, Issue 2 https://doi.org/10.1007/BF01024638	journal	February 1988
Alcohol production by Clostridium acetobutylicum induced by methyl viologen Rao, Govind; Mutharasan, R. Biotechnology Letters, Vol. 8, Issue 12 https://doi.org/10.1007/BF01078655	journal	December 1986
Iron effect on acetone-butanol fermentation Junelles, A. M.; Janati-Idrissi, R.; Petitdemange, H. Current Microbiology, Vol. 17, Issue 5 https://doi.org/10.1007/BF01571332	journal	September 1988
Carbon monoxide gasing leads to alcohol production and butyrate uptake without acetone formation in continuous cultures ofClostridium acetobutylicum Meyer, Charles L.; Roos, Joseph W.; Papoutsakis, Eleftherios T. Applied Microbiology and Biotechnology, Vol. 24, Issue 2 https://doi.org/10.1007/BF01982561	journal	May 1986
Achievements and perspectives to overcome the poor solvent resistance in acetone and butanol-producing microorganisms Ezeji, Thaddeus; Milne, Caroline; Price, Nathan D. Applied Microbiology and Biotechnology, Vol. 85, Issue 6 https://doi.org/10.1007/s00253-009-2390-0	journal	December 2009
A proteomic and transcriptional view of acidogenic and solventogenic steady-state cells of Clostridium acetobutylicum in a chemostat culture Janssen, Holger; Döring, Christina; Ehrenreich, Armin Applied Microbiology and Biotechnology, Vol. 87, Issue 6 https://doi.org/10.1007/s00253-010-2741-x	journal	July 2010
Controlling the oxidoreduction potential of the culture of Clostridium acetobutylicum leads to an earlier initiation of solventogenesis, thus increasing solvent productivity Wang, Shaohua; Zhu, Yan; Zhang, Yanping Applied Microbiology and Biotechnology, Vol. 93, Issue 3 https://doi.org/10.1007/s00253-011-3570-2	journal	September 2011
Effects of different replicons in conjugative plasmids on transformation efficiency, plasmid stability, gene expression and n-butanol biosynthesis in Clostridium tyrobutyricum Yu, Mingrui; Du, Yinming; Jiang, Wenyan Applied Microbiology and Biotechnology, Vol. 93, Issue 2 https://doi.org/10.1007/s00253-011-3736-y	journal	December 2011
Application of new metabolic engineering tools for Clostridium acetobutylicum Lütke-Eversloh, Tina Applied Microbiology and Biotechnology, Vol. 98, Issue 13 https://doi.org/10.1007/s00253-014-5785-5	journal	May 2014
The physical and catalytic properties of hydrogenase II of Clostridium pasteurianum. A comparison with hydrogenase I. Adams, M. W.; Mortenson, L. E. Journal of Biological Chemistry, Vol. 259, Issue 11 https://doi.org/10.1016/S0021-9258(17)39835-6	journal	June 1984
Regulation of solvent production in Clostridium acetobutylicum Girbal, Laurence; Soucaille, Philippe Trends in Biotechnology, Vol. 16, Issue 1 https://doi.org/10.1016/S0167-7799(97)01141-4	journal	January 1998
Developments in biobutanol production: New insights Kumar, Manish; Gayen, Kalyan Applied Energy, Vol. 88, Issue 6 https://doi.org/10.1016/j.apenergy.2010.12.055	journal	June 2011
The effects of pH on carbon material and energy balances in hydrogen-producing Clostridium tyrobutyricum JM1 Jo, Ji Hye; Lee, Dae Sung; Park, Jong Moon Bioresource Technology, Vol. 99, Issue 17 https://doi.org/10.1016/j.biortech.2008.03.060	journal	November 2008
Fed-batch fermentation for n-butanol production from cassava bagasse hydrolysate in a fibrous bed bioreactor with continuous gas stripping Lu, Congcong; Zhao, Jingbo; Yang, Shang-Tian Bioresource Technology, Vol. 104 https://doi.org/10.1016/j.biortech.2011.10.089	journal	January 2012
Refactoring redox cofactor regeneration for high-yield biocatalysis of glucose to butyric acid in Escherichia coli Lim, Jae Hyung; Seo, Sang Woo; Kim, Se Yeon Bioresource Technology, Vol. 135 https://doi.org/10.1016/j.biortech.2012.09.091	journal	May 2013
Butanol production from wood pulping hydrolysate in an integrated fermentation–gas stripping process Lu, Congcong; Dong, Jie; Yang, Shang-Tian Bioresource Technology, Vol. 143 https://doi.org/10.1016/j.biortech.2013.06.012	journal	September 2013
Stable high-titer n-butanol production from sucrose and sugarcane juice by Clostridium acetobutylicum JB200 in repeated batch fermentations Jiang, Wenyan; Zhao, Jingbo; Wang, Zhongqiang Bioresource Technology, Vol. 163 https://doi.org/10.1016/j.biortech.2014.04.047	journal	July 2014
Redox potential control and applications in microaerobic and anaerobic fermentations Liu, Chen-Guang; Xue, Chuang; Lin, Yen-Han Biotechnology Advances, Vol. 31, Issue 2 https://doi.org/10.1016/j.biotechadv.2012.11.005	journal	March 2013
Prospective and development of butanol as an advanced biofuel Xue, Chuang; Zhao, Xin-Qing; Liu, Chen-Guang Biotechnology Advances, Vol. 31, Issue 8 https://doi.org/10.1016/j.biotechadv.2013.08.004	journal	December 2013
Engineering solventogenic clostridia Papoutsakis, Eleftherios T. Current Opinion in Biotechnology, Vol. 19, Issue 5, p. 420-429 https://doi.org/10.1016/j.copbio.2008.08.003	journal	October 2008
Metabolic engineering of Clostridium acetobutylicum: recent advances to improve butanol production Lütke-Eversloh, Tina; Bahl, Hubert Current Opinion in Biotechnology, Vol. 22, Issue 5 https://doi.org/10.1016/j.copbio.2011.01.011	journal	October 2011
Cofactor engineering for advancing chemical biotechnology Wang, Yipeng; San, Ka-Yiu; Bennett, George N. Current Opinion in Biotechnology, Vol. 24, Issue 6 https://doi.org/10.1016/j.copbio.2013.03.022	journal	December 2013
Effect of pH on metabolic pathway shift in fermentation of xylose by Clostridium tyrobutyricum Zhu, Ying; Yang, Shang-Tian Journal of Biotechnology, Vol. 110, Issue 2 https://doi.org/10.1016/j.jbiotec.2004.02.006	journal	May 2004
Metabolic engineering of Clostridium tyrobutyricum for n-butanol production Yu, Mingrui; Zhang, Yali; Tang, I-Ching Metabolic Engineering, Vol. 13, Issue 4 https://doi.org/10.1016/j.ymben.2011.04.002	journal	July 2011
Activating transhydrogenase and NAD kinase in combination for improving isobutanol production Shi, Aiqin; Zhu, Xinna; Lu, Jiao Metabolic Engineering, Vol. 16 https://doi.org/10.1016/j.ymben.2012.11.008	journal	March 2013
Model-driven rebalancing of the intracellular redox state for optimization of a heterologous n-butanol pathway in Escherichia coli Lim, Jae Hyung; Seo, Sang Woo; Kim, Se Yeon Metabolic Engineering, Vol. 20 https://doi.org/10.1016/j.ymben.2013.09.003	journal	November 2013
Construction and Characterization of ack Deleted Mutant of Clostridium tyrobutyricum for Enhanced Butyric Acid and Hydrogen Production Liu, Xiaoguang; Zhu, Ying; Yang, Shang-Tian Biotechnology Progress, Vol. 22, Issue 5 https://doi.org/10.1021/bp060082g	journal	January 2006
Enzyme mechanism as a kinetic control element for designing synthetic biofuel pathways Bond-Watts, Brooks B.; Bellerose, Robert J.; Chang, Michelle C. Y. Nature Chemical Biology, Vol. 7, Issue 4 https://doi.org/10.1038/nchembio.537	journal	February 2011
How neutral red modified carbon and electron flow in Clostridium acetobutylicum grown in chemostat culture at neutral pH Girbal, Laurence; Vasconcelos, Isabel; Saint-Amans, Silvie FEMS Microbiology Reviews, Vol. 16, Issue 2-3 https://doi.org/10.1111/j.1574-6976.1995.tb00163.x	journal	February 1995
Driving Forces Enable High-Titer Anaerobic 1-Butanol Synthesis in Escherichia coli Shen, Claire R.; Lan, Ethan I.; Dekishima, Yasumasa Applied and Environmental Microbiology, Vol. 77, Issue 9, p. 2905-2915 https://doi.org/10.1128/AEM.03034-10	journal	March 2011
Altered Electron Flow in Continuous Cultures of Clostridium acetobutylicum Induced by Viologen Dyes Rao, Govind; Mutharasan, R. Applied and Environmental Microbiology, Vol. 53, Issue 6 https://doi.org/10.1128/AEM.53.6.1232-1235.1987	journal	January 1987
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 Fontaine, L.; Meynial-Salles, I.; Girbal, L. Journal of Bacteriology, Vol. 184, Issue 3 https://doi.org/10.1128/JB.184.3.821-830.2002	journal	February 2002
Acetone-butanol fermentation revisited. Jones, D. T.; Woods, D. R. Microbiological Reviews, Vol. 50, Issue 4 https://doi.org/10.1128/MMBR.50.4.484-524.1986	journal	January 1986
Regulation of carbon and electron flow in Clostridium acetobutylicum grown in chemostat culture at neutral pH on mixtures of glucose and glycerol. Vasconcelos, I.; Girbal, L.; Soucaille, P. Journal of Bacteriology, Vol. 176, Issue 5 https://doi.org/10.1128/jb.176.5.1443-1450.1994	journal	March 1994
A systems biology approach to investigate the effect of pH-induced gene regulation on solvent production by Clostridium acetobutylicum in continuous culture Haus, Sylvia; Jabbari, Sara; Millat, Thomas BMC Systems Biology, Vol. 5, Issue 1 https://doi.org/10.1186/1752-0509-5-10	journal	January 2011
Continuous Production of Butanol by Clostridium acetobutylicum Immobilized in a Fibrous Bed Bioreactor Huang, Wei-Cho; Ramey, David E.; Yang, Shang-Tian Applied Biochemistry and Biotechnology, Vol. 115, Issue 1-3 https://doi.org/10.1385/ABAB:115:1-3:0887	journal	January 2004
Metabolic Process Engineering for Biochemicals and Biofuels Production Yang, Shang-Tian Journal of Microbial & Biochemical Technology, Vol. 06, Issue 02 https://doi.org/10.4172/1948-5948.1000e116	journal	January 2014

Similar Records

Engineering Clostridium cellulovorans for highly selective n ‐butanol production from cellulose in consolidated bioprocessing

Journal Article · Thu Apr 22 20:00:00 EDT 2021 · Biotechnology and Bioengineering · OSTI ID:1780198

Characterization of a Clostridium beijerinckii spo0A mutant and its application for butyl butyrate production

Journal Article · Tue Aug 16 20:00:00 EDT 2016 · Biotechnology and Bioengineering · OSTI ID:1401466

Production of a Biofuel Intermediate from Biomass Sugars by Clostridium tyrobutyricum

Conference · Thu Dec 05 23:00:00 EST 2019 · OSTI ID:1578266

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

Citation Formats

References (45)

Similar Records

Related Subjects