Systems metabolic engineering of Corynebacterium glutamicum for efficient l-tryptophan production

<i>Corynebacterium glutamicum</i> is a versatile industrial microorganism for producing various amino acids. However, there have been no reports of well-defined <i>C. glutamicum</i> strains capable of hyperproducing l-tryptophan. This study presents a comprehensive metabolic engineering approach to establish robust <i>C. glutamicum</i> strains for l-tryptophan biosynthesis, including: (1) identification of potential targets by enzyme-constrained genome-scale modeling; (2) enhancement of the l-tryptophan biosynthetic pathway; (3) reconfiguration of central metabolic pathways; (4) identification of metabolic bottlenecks through comparative metabolome analysis; (5) engineering of the transport system, shikimate pathway, and precursor supply; and (6) repression of competing pathways and iterative optimization of key targets. The resulting <i>C. glutamicum</i> strain achieved a remarkable l-tryptophan titer of 50.5 g/L in 48h with a yield of 0.17 g/g glucose in fed-batch fermentation. This study highlights the efficacy of integrating computational modeling with systems metabolic engineering for significantly enhancing the production capabilities of industrial microorganisms.