Technical guide for genetic advancement of underdeveloped and intractable Clostridium
Abstract
In recent years, the genus Clostridium has risen to the forefront of both medical biotechnology and industrial biotechnology owing to its potential in applications as diverse as anticancer therapy and production of commodity chemicals and biofuels. The prevalence of hyper-virulent strains of C. difficile within medical institutions has also led to a global epidemic that demands a more thorough understanding of clostridial genetics, physiology, and pathogenicity. Unfortunately, Clostridium suffers from a lack of sophisticated genetic tools and techniques which has hindered the biotechnological exploitation of this important bacterial genus. This review provides a comprehensive summary of biotechnological progress made in clostridial genetic tool development, while also aiming to serve as a technical guide for the advancement of underdeveloped clostridial strains, including recalcitrant species, novel environmental samples, and non-type strains. Relevant strain engineering techniques, from genome sequencing and establishment of a gene transfer methodology through to deployment of advanced genome editing procedures, are discussed in detail to provide a blueprint for future clostridial strain construction endeavors. It is expected that a more thorough and rounded-out genetic toolkit available for use in the clostridia will bring about the construction of superior bioprocessing strains and a more complete understanding of clostridial genetics, physiology, and pathogenicity.
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DNA transfer is especially challenging for Gram-positive bacteria such as clostridia and most other acetogens due to the physical barrier of the thick peptidoglycan cell wall and highly active restriction-modification systems.
“This is especially true for Gram-positive bacteria such as clostridia and most other acetogens due to the physical barrier of the thick peptidoglycan cell wall and highly active restriction-modification systems (Pyne et al., 2014).”
From Gas Fermentation—A Flexible Platform for Commercial Scale Production of Low-Carbon-Fuels and Chemicals from Waste and Renewable Feedstocks (2016) · cites this paper as Pyne et al., 2014
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Liew, F., Martin, M. E., Tappel, R. C., Heijstra, B. D., Mihalcea, C., & Köpke, M. (2016) · Frontiers in Microbiology · doi:10.3389/fmicb.2016.00694 · Cited by 485 · see citing works
Christi, K., Hudson, J., & Egan, S. (2024) · Microbiological Research · doi:10.1016/j.micres.2024.127729 · Cited by 8 · see citing works
- Authors
- Pyne, M. E., Bruder, M., Moo-Young, M., Chung, D. A., & Chou, C. P.
- Affiliations
- University of Waterloo (CA)
- Corresponding author
- Chung, Duane A.
- Venue
- Biotechnology Advances
- Publisher
- Elsevier BV
- Published
- 2014-05
- Volume
- 32
- Issue
- 3
- Pages
- 623-641
- ISSN
- 0734-9750 (ISSN-L)
- OpenAlex
- W2106986485
- Cited by
- 78
- References
- 198
- Citations by year
- 2025: 4 · 2024: 5 · 2023: 4 · 2022: 2 · 2021: 6 · 2020: 13 · 2019: 14 · 2018: 6 · 2017: 2 · 2016: 10 · 2015: 7 · 2014: 4
- FWCI
- 0.72
- Type
- journal article
- Language
- en
- Open access
- closed
- Locations
- Biotechnology Advances (publishedVersion)
- PubMed (publishedVersion)
- Funders
- Natural Sciences and Engineering Research Council of Canada
- Canada Research Chairs
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Microbial Metabolic Engineering and Bioproduction
- Health Sciences › Medicine › Infectious Diseases › Clostridium difficile and Clostridium perfringens research
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Genetics › Bacterial Genetics and Biotechnology
- Keywords
- Clostridium, genetic instruments, genome editing, gene transfer, strain engineering, Clostridium difficile, hypervirulent strains, genome sequencing, industrial biotechnology, medical biotechnology
- MeSH terms
- Clostridium, Genetic Engineering, Industrial Microbiology, Genome, Bacterial, Biofuels
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