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Deciphering of the Mannitol Metabolism Pathway in Clostridium tyrobutyricum ATCC 25755 by Comparative Transcriptome Analysis

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Abstract

Clostridium tyrobutyricum has great potential for bio-based chemicals and biofuel production from mannitol; however, the mannitol metabolic pathway and its metabolic regulatory mechanism have not been elucidated. To this end, the RNA-seq analysis on the mid-log growth phase of C. tyrobutyricum grown on mannitol or xylose was performed. Comparative transcriptome analysis and co-transcription experiment indicated that mtlARFD, which encodes the mannitol-specific IIA component, transcription activator, mannitol-specific IIBC components, and mannitol-1-phosphate 5-dehydrogenase, respectively, formed a polycistronic operon and could be responsible for mannitol uptake and metabolism. In addition, comparative genomic analysis of the mtlARFD organization and the MtlR protein structural domain among various Firmicutes strains identified the putative cre (catabolite-responsive element) sites and conserved phosphorylation sites, but whether the expression of mannitol operon was affected by CcpA- and MtlR-mediated metabolic regulation during mixed substrate fermentation needs to be further verified experimentally. Based on the gene knockout and complementation results, the predicted mannitol operon mtlARFD was confirmed to be responsible for mannitol utilization in C. tyrobutyricum. The results of this study could be used to enhance the mannitol metabolic pathway and explore the potential metabolic regulation mechanism of mannitol during mixed substrate fermentation.

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Availability of Data and Materials

The data sets generated and/or analyzed during the current study are available from the corresponding author on reasonable request.

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Funding

This work was supported by National Natural Science Foundation of China (21808069 and 22178133), Guangzhou Basic and Applied Basic Research (202102020281), and Science and Technology Planning Project of Guangdong Province of China (2019A050510008).

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Authors and Affiliations

  1. School of Biology and Biological Engineering, South China University of Technology, Guangzhou, 510006, China

    Hongxin Fu, Lu Yang, Huihui Zhang & Jufang Wang

  2. Guangdong Key Laboratory of Fermentation and Enzyme Engineering, South China University of Technology, Guangzhou, 510006, China

    Hongxin Fu & Jufang Wang

  3. State Key Laboratory of Pulp and Paper Engineering, South China University of Technology, Guangzhou, 510006, China

    Hongxin Fu & Jufang Wang

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  1. Hongxin Fu
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  2. Lu Yang
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Contributions

HXF and JFW conceived and designed research. LY and HHZ conducted experiments. HXF and LY analyzed data. HXF wrote the manuscript. All authors read and approved the manuscript.

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Correspondence to Hongxin Fu or Jufang Wang.

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Fu, H., Yang, L., Zhang, H. et al. Deciphering of the Mannitol Metabolism Pathway in Clostridium tyrobutyricum ATCC 25755 by Comparative Transcriptome Analysis. Appl Biochem Biotechnol 195, 1072–1084 (2023). https://doi.org/10.1007/s12010-022-04209-8

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