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A novel regulatory pathway consisting of a two-component system and an ABC-type transporter contributes to butanol tolerance in Clostridium acetobutylicum

  • Applied microbial and cell physiology
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Abstract

Despite the long-term interest in solventogenic clostridia-based ABE (acetone-butanol-ethanol) fermentation, clostridial butanol tolerance and its underlying mechanism remain poorly understood, which is a major obstacle hindering further improvements of this important fermentative process. In this study, a two-component system (TCS), BtrK/BtrR, was identified and demonstrated to positively regulate butanol tolerance and ABE solvent formation in Clostridium acetobutylicum, a representative species of solventogenic clostridia. The transcriptomic analysis results showed that BtrK/BtrR has a pleiotropic regulatory function, affecting a large number of crucial genes and metabolic pathways. Of the differentially expressed genes, btrTM, encoding a putative ABC-type transporter (named BtrTM), was shown to be under the direct control of BtrR, the response regulator of the BtrK/BtrR TCS. Furthermore, BtrTM was shown to contribute to more butanol tolerance (46.5% increase) by overexpression, revealing a novel regulatory mechanism consisting of the BtrK/BtrR TCS and the BtrTM transporter in C. acetobutylicum. Based on these findings, we achieved faster growth and solvent production of C. acetobutylicum by overexpressing BtrK/BtrR or its direct target BtrTM, although no significant improvement in the final butanol titer and yield. These results further confirm the importance of BtrK/BtrR and BtrTM in this organism. Also, of significance, a specific number of btrR-btrT-btrM-btrK-like gene clusters were identified in other Clostridium species, including the pathogens Clostridium perfringens and Clostridium botulinum, indicating a broad role for this regulatory module in the class Clostridia.

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Funding

This work was supported by the National Natural Science Foundation of China (31630003 and 31921006), and Open Funding Project of the State Key Laboratory of Bioreactor Engineering.

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  1. Yunpeng Yang, Nannan Lang and Lu Zhang contributed equally to this work.

Authors and Affiliations

  1. Key Laboratory of Synthetic Biology, CAS Center for Excellence in Molecular Plant Sciences, Shanghai Institute of Plant Physiology and Ecology, Chinese Academy of Sciences, 300 Fenglin Road, Shanghai, 200032, China

    Yunpeng Yang, Nannan Lang, Lu Zhang, Weihong Jiang & Yang Gu

  2. State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology, Shanghai, 200237, China

    Hui Wu

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  1. Yunpeng Yang
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  2. Nannan Lang
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  3. Lu Zhang
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Contributions

W. J. and Y. G. designed the research; Y. Y., N. L., and L. Z. performed the research; H. W. analyzed the data; and Y.Y., W. J., and Y. G. wrote the paper.

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Correspondence to Weihong Jiang or Yang Gu.

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Yang, Y., Lang, N., Zhang, L. et al. A novel regulatory pathway consisting of a two-component system and an ABC-type transporter contributes to butanol tolerance in Clostridium acetobutylicum. Appl Microbiol Biotechnol 104, 5011–5023 (2020). https://doi.org/10.1007/s00253-020-10555-6

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