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Biotechnology

Driving Forces Enable High-Titer Anaerobic 1-Butanol Synthesis in Escherichia coli

Claire R. Shen, Ethan I. Lan, Yasumasa Dekishima, Antonino Baez, Kwang Myung Cho, James C. Liao
Claire R. Shen
1Department of Chemical and Biomolecular Engineering, University of California, Los Angeles, California
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Ethan I. Lan
1Department of Chemical and Biomolecular Engineering, University of California, Los Angeles, California
2Biomedical Engineering Interdepartmental Program, University of California, Los Angeles, California
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Yasumasa Dekishima
1Department of Chemical and Biomolecular Engineering, University of California, Los Angeles, California
4Mitsubishi Chemical Group Science and Technology Research Center, Inc., Yokohama, Japan
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Antonino Baez
1Department of Chemical and Biomolecular Engineering, University of California, Los Angeles, California
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Kwang Myung Cho
1Department of Chemical and Biomolecular Engineering, University of California, Los Angeles, California
3Institute for Genomics and Proteomics, University of California, Los Angeles, California
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James C. Liao
1Department of Chemical and Biomolecular Engineering, University of California, Los Angeles, California
2Biomedical Engineering Interdepartmental Program, University of California, Los Angeles, California
3Institute for Genomics and Proteomics, University of California, Los Angeles, California
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  • For correspondence: liaoj@ucla.edu
DOI: 10.1128/AEM.03034-10
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ABSTRACT

1-Butanol, an important chemical feedstock and advanced biofuel, is produced by Clostridium species. Various efforts have been made to transfer the clostridial 1-butanol pathway into other microorganisms. However, in contrast to similar compounds, only limited titers of 1-butanol were attained. In this work, we constructed a modified clostridial 1-butanol pathway in Escherichia coli to provide an irreversible reaction catalyzed by trans-enoyl-coenzyme A (CoA) reductase (Ter) and created NADH and acetyl-CoA driving forces to direct the flux. We achieved high-titer (30 g/liter) and high-yield (70 to 88% of the theoretical) production of 1-butanol anaerobically, comparable to or exceeding the levels demonstrated by native producers. Without the NADH and acetyl-CoA driving forces, the Ter reaction alone only achieved about 1/10 the level of production. The engineered host platform also enables the selection of essential enzymes with better catalytic efficiency or expression by anaerobic growth rescue. These results demonstrate the importance of driving forces in the efficient production of nonnative products.

  • Copyright © 2011, American Society for Microbiology

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Driving Forces Enable High-Titer Anaerobic 1-Butanol Synthesis in Escherichia coli
Claire R. Shen, Ethan I. Lan, Yasumasa Dekishima, Antonino Baez, Kwang Myung Cho, James C. Liao
Applied and Environmental Microbiology Apr 2011, 77 (9) 2905-2915; DOI: 10.1128/AEM.03034-10

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Driving Forces Enable High-Titer Anaerobic 1-Butanol Synthesis in Escherichia coli
Claire R. Shen, Ethan I. Lan, Yasumasa Dekishima, Antonino Baez, Kwang Myung Cho, James C. Liao
Applied and Environmental Microbiology Apr 2011, 77 (9) 2905-2915; DOI: 10.1128/AEM.03034-10
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