Transcriptome and metabolome analyses reveal novel genetic targets for L-tryptophan overproduction in Corynebacterium glutamicum

Abstract Corynebacterium glutamicum is a promising microbial chassis for the industrial production of L-tryptophan, which has exhibited increasing demand due to its diverse applications and high market value. In previous work, we developed an L-tryptophan-overproducing C. glutamicum strain TR26 through multiple rounds of rational metabolic engineering. Here, comparative transcriptome and metabolome analyses were conducted between TR26 and its progenitor strain MB001 to reveal the underlying mechanisms and potential bottlenecks for L-tryptophan production in TR26. Furthermore, by systematically down- and up-regulating differentially expressed genes of interest, two novel genetic targets, glnK and sugR , were identified as being associated with L-tryptophan synthesis. Specifically, the repression of glnK and overexpression of sugR in strain TR26 increased the titer of L-tryptophan by 6.7% and 20.9%, respectively. Gene transcription profiling and intracellular metabolite analysis further suggested that the observed improvements in L-tryptophan synthesis could be attributed to optimized nitrogen transport and metabolism, efficient reallocation of cellular resources and enhanced supply of phosphoenolpyruvate (PEP). This study advances our understanding of the regulation mechanisms governing L-tryptophan synthesis in C. glutamicum and provides valuable insights for further optimization of industrial cell factories.