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Solventogenic clostridia lose their solvent formation capabilities during repeated sub-culturing of batch cultures or during a continuous fermentation. This phenomenon is known as ‘degeneration’ and is one of the limiting factors to maximize butanol production. Therefore, it is needed to detect and monitor degenerated solventogenic clostridia for high butanol production without failure
of operation. In this study, we designed specific primers and probe sets for real-time PCR to detect the degenerated Clostridium acetobutylicum using sequences from the pSOL1 plasmid. Degeneration of Clostridium acetobutylicum is associated with a loss of pSOL1, which carries all the genes involved in the final steps of solvent formation. We confirmed that these pSOL1 sequence based primer and probe set could be used to detect non-degenerated Clostridium
acetobutyricum specifically which posses pSOL1. Monitoring and quantification of degenerated Clostridium acetobutylicum, which lost their pSOL1 plasmid, were also investigated during repeated sub-culturing of batch culture by using real-time qPCR.
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