Non-natural 2-methyl-1-butanol (2 MB) has been biosynthesized through the modification of metabolic pathways using Corynebacterium crenatum, a non-model host. However, its production capacity is not effectively improved. In this study, the fermentation process was strengthened through factor combination design (FCD) for enhancing the production of 2 MB. Our results showed that the highest production of 2 MB, 3-methyl-1-butanol (3 MB), ethanol, and total solvent was 4.87 ± 0.39 g/L, 3.57 ± 0.21 g/L, 5.74 ± 0.43 g/L, and 14.18 g/L, respectively, under the optimal fermentation conditions. The optimal fermentation conditions were determined through the FCD to be as follows: pH of 6.5, IPTG concentration of 1.2 mM, fermentation temperature of 32 °C, and fermentation time of 96 h. This study provides a significant guidance for the optimal control technology of the genetically engineered C. crenatum, and also a useful reference for the industrial production of 2 MB via the microbial fermentation approach.