Comparative transcriptome analysis of Clostridium tyrobutyricum expressing a heterologous uptake hydrogenase
2020
Li, Weiming | Cheng, Chi | Cao, Guangli | Yang, Shang-Tian | Ren, Nanqi
Clostridium tyrobutyricum is a promising microbial cell factory to produce biofuels. In this study, an uptake hydrogenase (hyd2293) from Ethanoligenens harbinense was overexpressed in C. tyrobutyricum and significantly affected the redox reactions and metabolic profiles. Compared to the parental strain (Ct-WT), the mutant strain Ct-Hyd2293 produced ~34% less butyrate, ~148% more acetate, and ~11% less hydrogen, accompanied by the emerging genesis of butanol. Comparative transcriptome analysis revealed that 666 genes were significantly differentially expressed after the overexpression of hyd2293, including 82 up-regulated genes and 584 down-regulated genes. The up-regulated genes were mainly involved in carbohydrate and energy metabolisms while the down-regulated genes were distributed in nearly all pathways. Genes involved in glucose transportation, glycolysis, different fermentation pathways and hydrogen metabolism were studied and the gene expression changes showed the mechanism of the metabolic flux redistribution in Ct-Hyd2293. The overexpression of uptake hydrogenase redirected electrons from hydrogen and butyrate to butanol. The key enzymes participating in the energy conservation and sporulation were also identified and their transcription levels were generally reduced. This study demonstrated the transcriptomic responses of C. tyrobutyricum to the expression of a heterologous uptake hydrogenase, which provided a better understanding of the metabolic characteristics of C. tyrobutyricum and demonstrated the potential role of redox manipulation in metabolic engineering for biofuel productions.
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