2019•Microbial BiotechnologyOpen access

Control of solvent production by sigma‐54 factor and the transcriptional activator AdhR in Clostridium beijerinckii

Bin Yang, Xiaoqun Nie, Yang Gu, Weihong Jiang, Chen Yang

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Abstract

Summary Clostridia are obligate anaerobic bacteria that can produce solvents such as acetone, butanol and ethanol. Alcohol dehydrogenases (ADHs) play a key role in solvent production; however, their regulatory mechanisms remain largely unknown. In this study, we characterized the regulatory mechanisms of two ADH‐encoding genes in C. beijerinckii . SigL (sigma factor σ 54 ) was found to be required for transcription of adhA1 and adhA2 genes. Moreover, a novel transcriptional activator AdhR was identified, which binds to the σ 54 promoter and activates σ 54 ‐dependent transcription of adhA1 and adhA2 . Clostridia beijerinckii mutants deficient in SigL or AdhR showed severely impaired butanol and ethanol production as well as altered acetone and butyrate synthesis. Overexpression of SigL resulted in significantly improved solvent production by C. beijerinckii when butyrate was added to cultures. Our results reveal SigL as a novel engineering target for improving solvent production by C. beijerinckii and other solvent‐producing clostridia. Moreover, this study gains an insight into regulation of alcohol metabolism in diverse clostridia.

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Summary Clostridia are obligate anaerobic bacteria that can produce solvents such as acetone, butanol and ethanol. Alcohol dehydrogenases (ADHs) play a key role in solvent production; however, their regulatory mechanisms remain largely unknown. In this study, we characterized the regulatory mechanisms of two ADH‐encoding genes in C. beijerinckii . SigL (sigma factor σ 54 ) was found to be required for transcription of adhA1 and adhA2 genes. Moreover, a novel transcriptional activator AdhR was identified, which binds to the σ 54 promoter and activates σ 54 ‐dependent transcription of adhA1 and adhA2 . Clostridia beijerinckii mutants deficient in SigL or AdhR showed severely impaired butanol and ethanol production as well as altered acetone and butyrate synthesis. Overexpression of SigL resulted in significantly improved solvent production by C. beijerinckii when butyrate was added to cultures. Our results reveal SigL as a novel engineering target for improving solvent production by C. beijerinckii and other solvent‐producing clostridia. Moreover, this study gains an insight into regulation of alcohol metabolism in diverse clostridia.

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Available abstract

Summary Clostridia are obligate anaerobic bacteria that can produce solvents such as acetone, butanol and ethanol. Alcohol dehydrogenases (ADHs) play a key role in solvent production; however, their regulatory mechanisms remain largely unknown. In this study, we characterized the regulatory mechanisms of two ADH‐encoding genes in C. beijerinckii . SigL (sigma factor σ 54 ) was found to be required for transcription of adhA1 and adhA2 genes. Moreover, a novel transcriptional activator AdhR was identified, which binds to the σ 54 promoter and activates σ 54 ‐dependent transcription of adhA1 and adhA2 . Clostridia beijerinckii mutants deficient in SigL or AdhR showed severely impaired butanol and ethanol production as well as altered acetone and butyrate synthesis. Overexpression of SigL resulted in significantly improved solvent production by C. beijerinckii when butyrate was added to cultures. Our results reveal SigL as a novel engineering target for improving solvent production by C. beijerinckii and other solvent‐producing clostridia. Moreover, this study gains an insight into regulation of alcohol metabolism in diverse clostridia.

Key concepts: Clostridium beijerinckii, Clostridia, Butanol, Chemistry, Clostridium, Sigma factor, Alcohol dehydrogenase, Biochemistry

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