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The solventogenic  C .  beijerinckii  DSM 6423, a microorganism that naturally produces isopropanol and butanol, was previously modified by random mutagenesis. In this work, one of the resulting mutants was characterized. This strain, selected with allyl alcohol and designated as the AA mutant, shows a dominant production of acids, a severely diminished butanol synthesis capacity, and produces acetone instead of isopropanol. Interestingly, this solvent-deficient strain was also found to have a limited consumption of two carbohydrates and to be still able to form spores, highlighting its particular phenotype. Sequencing of the AA mutant revealed point mutations in several genes including CIBE_0767 ( sigL ), which encodes the σ 54  sigma factor. Complementation with wild-type  sigL  fully restored solvent production and sugar assimilation and RT-qPCR analyses revealed its transcriptional control of several genes related to solventogensis, demonstrating the central role of σ 54  in  C .  beijerinckii  DSM 6423. Comparative genomics analysis suggested that this function is conserved at the species level, and this hypothesis was further confirmed through the deletion of  sigL  in the model strain  C .  beijerinckii  NCIMB 8052.

σ54 (σL) plays a central role in carbon metabolism in the industrially relevant Clostridium beijerinckii