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The Butanol Producing Microbe Clostridium beijerinckii NCIMB 14988 Manipulated Using Forward and Reverse Genetic
Gareth T Little1, Benjamin J Willson1, John T Heap1
1Clostridia Research Group, BBSRC/EPSRC Synthetic Biology Research Centre (SBRC), University of Nottingham, University Park, Nottingham NG7 2RD, UK.
Biotechnology Journal
|April 17, 2018
Summary
We characterized Clostridium beijerinckii NCIMB 14988, a novel strain for sustainable solvent production. We developed genetic tools, including gene knockout and point mutation techniques, enabling further metabolic engineering for biofuel applications.
Area of Science:
- Microbiology
- Synthetic Biology
- Biotechnology
Background:
- Clostridium beijerinckii has potential for sustainable bioconversion of carbohydrates into solvents.
- Limited genomic characterization and genetic tools exist for many Clostridium strains.
Purpose of the Study:
- To characterize a new strain, Clostridium beijerinckii NCIMB 14988, for biofuel production.
- To develop and demonstrate genetic tools for this strain.
Main Methods:
- Genome sequencing of C. beijerinckii NCIMB 14988.
- Development of a high-efficiency plasmid transformation protocol.
- Application of transposon mutagenesis, ClosTron, and allelic exchange for gene knockout and point mutation.
Main Results:
- Complete genome sequence of C. beijerinckii NCIMB 14988 determined.
- Efficient genetic manipulation techniques established, including gene inactivation and point mutations.
- Disruption of a non-coding region resulted in a non-sporulating phenotype.
Conclusions:
- C. beijerinckii NCIMB 14988 is a promising strain for sustainable solvent production.
- The developed genetic toolkit facilitates metabolic engineering and further characterization of this strain.
- Genetic manipulation can impact key cellular processes like sporulation.
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