Abstract:[Objective] This study aimed to investigate the feasibility of the fermentation process for L-lysine production by immobilized Corynebacterium glutamicum.[Method] Firstly, the effects of different volume fractions of seed medium on the fermentation stability of immobilized Corynebacterium glutamicum were examined. Subsequently, the impacts of immobilization carriers with different specifications on the fermentation performance were analyzed. Finally, a comparative analysis was conducted on the differences between the fermentation processes with free and immobilized Corynebacterium glutamicum.[Result] The addition of 5% volume fraction of seed medium adequately sustained the growth and metabolic activities of Corynebacterium glutamicum. Higher volume fractions diverted carbon sources towards bacterial growth, thereby limiting product synthesis. The most favorable fermentation metrics were achieved with a specification of 0.061 g/cm2. Compared with free Corynebacterium glutamicum fermentation, immobilized Corynebacterium glutamicum exhibited a significantly shortened fermentation cycle and an increase of 16.24% in the acid production rate, which was attributable to the high cell density and protective effects of immobilization. Furthermore, the reduced biomass redirected more carbon sources towards product synthesis, resulting in higher conversion rates.[Conclusion] The novel fermentation process employing immobilized Corynebacterium glutamicum demonstrates strong application feasibility, providing important insights for its future industrial implementation.