The Butanol Producing Microbe Clostridium beijerinckii NCIMB 14988 Manipulated Using Forward and Reverse Genetic Tools

• DOI: 10.1002/biot.201700711
• Publication Date: 2018-04-16T21:11:25Z
• Authors: Gareth T. Little; Benjamin J. Willson; John T. Heap; Klaus Winzer; Nigel P. Minton
• Keywords: Clostridium beijerinckii; Pentose; Group II intron; Clostridia

The solventogenic anaerobe Clostridium beijerinckii has potential for use in the sustainable bioconversion of plant‐derived carbohydrates into solvents, such as butanol or acetone. However, relatively few strains have been extensively characterised either at genomic level through exemplification a complete genetic toolkit. To remedy this situation, new strain C. , NCIMB 14988, is selected from among total 55 clostridial isolates capable growth on hexose and pentose sugars. Chosen basis its favorable properties, genome sequence 14988 determined high‐efficiency plasmid transformation protocol devised. developed DNA transfer procedure allowed demonstration forward reverse techniques transposon mutagenesis gene knockout, respectively. latter accomplished successful deployment both group II intron retargeting (ClosTron) allelic exchange. In addition to inactivation, exchange used create point mutations chromosome, allowing effect amino acid changes enzymes involved primary metabolism be characterized. ClosTron mediated disruption currently unannotated non‐coding region between genes LF65_05915 LF65_05920 found result non‐sporulating phenotype.