The increasing depletion of traditional energy and the greenhouse effect pose challenges to global sustainable development. Butyl butyrate’s low viscosity and low carbon emission make it a biofuel with a high potential of alleviating the above problems. We genetically engineered C. tyrobutyricum to enable efficient and large-scale synthesis of butyl butyrate. We optimize metabolic pathways through three strategies to further reinforce the synthesis of butyrate and butanol, including precursor accumulation, inhibition of byproducts, and increasing mass transfer efficiency as well as promoter engineering. The results showed that yields of butyl butyrate can be improved to varying degrees, which indicated that it is a promising way to improve the biosynthesis efficiency of butyl butyrate.