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Gene Editing Technologies for Biofuel Production in Thermophilic Microbes

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Metabolic Pathway Engineering

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2096))

Abstract

Thermophilic microbes are an attractive bioproduction platform due to their inherently lower contamination risk and their ability to perform thermostable enzymatic processes which may be required for biomass processing and other industrial applications. The engineering of microbes for industrial scale processes requires a suite of genetic engineering tools to optimize existing biological systems as well as to design and incorporate new metabolic pathways within strains. Yet, such tools are often lacking and/or inadequate for novel microbes, especially thermophiles. This chapter focuses on genetic tool development and engineering strategies, in addition to challenges, for thermophilic microbes. We provide detailed instructions and techniques for tool development for an anaerobic thermophile, Caldanaerobacter subterraneus subsp. tengcongensis, including culturing, plasmid construction, transformation, and selection. This establishes a foundation for advanced genetic tool development necessary for the metabolic engineering of this microbe and potentially other thermophilic organisms.

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Acknowledgments

This work was supported by the Office of Energy Efficiency and Renewable Energy Bioenergy Technologies Office. This work was authored in part by Alliance for Sustainable Energy, LLC, the Manager and Operator of the National Renewable Energy Laboratory for the U.S. Department of Energy (DOE) under Contract No. DE-AC36-08GO28308. The views expressed in the chapter do not necessarily represent the views of the DOE or the U.S. Government. The U.S. Government retains and the publisher, by accepting the article for publication, acknowledges that the U.S. Government retains a nonexclusive, paid-up, irrevocable, worldwide license to publish or reproduce the published form of this work, or allow others to do so for U.S. Government purposes.

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Authors and Affiliations

  1. National Renewable Energy Laboratory, Golden, CO, USA

    Sharon Smolinski, Emily Freed & Carrie Eckert

  2. Renewable and Sustainable Energy Institute, University of Colorado, Boulder, CO, USA

    Emily Freed & Carrie Eckert

Authors
  1. Sharon Smolinski
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  2. Emily Freed
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  3. Carrie Eckert
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Correspondence to Carrie Eckert .

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Editors and Affiliations

  1. National Renewable Energy Laboratory, Golden, CO, USA

    Michael E. Himmel

  2. National Renewable Energy Laboratory, Golden, CO, USA

    Yannick J. Bomble

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Smolinski, S., Freed, E., Eckert, C. (2020). Gene Editing Technologies for Biofuel Production in Thermophilic Microbes. In: Himmel, M., Bomble, Y. (eds) Metabolic Pathway Engineering. Methods in Molecular Biology, vol 2096. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-0195-2_12

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  • DOI: https://doi.org/10.1007/978-1-0716-0195-2_12

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-0194-5

  • Online ISBN: 978-1-0716-0195-2

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