Chapter 12: Gene Editing Technologies for Biofuel Production in Thermophilic Microbes

Sharon Smolinski, Emily Freed, Carrie Eckert

Research output: Chapter in Book/Report/Conference proceedingChapterpeer-review

2 Scopus Citations


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.

Original languageAmerican English
Title of host publicationMetabolic Pathway Engineering
Subtitle of host publicationMethods in Molecular Biology, Volume 2096
EditorsM. E. Himmel, Y. J. Bomble
PublisherHumana Press Inc.
Number of pages15
StatePublished - 2020

Publication series

NameMethods in Molecular Biology
ISSN (Print)1064-3745
ISSN (Electronic)1940-6029

Bibliographical note

Publisher Copyright:
© Springer Science+Business Media, LLC, part of Springer Nature 2020.

NREL Publication Number

  • NREL/CH-2700-77588


  • Biofuel
  • Caldanaerobacter subterraneus subsp. tengcongensis
  • Genetic engineering
  • Genetic tools
  • Thermophile
  • Transformation


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