Technical guide for genetic advancement of underdeveloped and intractable Clostridium.
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.
Department of Chemical Engineering, University of Waterloo, Waterloo, Ontario N2L 3G1, Canada.
Michael E
ME
Pyne
Department of Chemical Engineering, University of Waterloo, Waterloo, Ontario N2L 3G1, Canada.
Mark
M
Bruder
Department of Chemical Engineering, University of Waterloo, Waterloo, Ontario N2L 3G1, Canada.
Murray
M
Moo-Young
Department of Chemical Engineering, University of Waterloo, Waterloo, Ontario N2L 3G1, Canada; Algaeneers Inc., Hamilton, Ontario L8N 3Z5, Canada. Electronic address: duane.chung@uwaterloo.ca.
Duane A
DA
Chung
Department of Chemical Engineering, University of Waterloo, Waterloo, Ontario N2L 3G1, Canada. Electronic address: cpchou@uwaterloo.ca.
C Perry
CP
Chou
D056804:Biofuels; D003013:Clostridium / Q000235:genetics / Q000502:physiology; D005818:Genetic Engineering; D016680:Genome, Bacterial / Q000235:genetics*; D007218:Industrial Microbiology
D056804:Biofuels
Biofuels; Clostridia; Clostridium; Gene knockout; Genetic engineering; Industrial biotechnology; Medical biotechnology; Overexpression; Transformation