β-carotene is an indispensable nutrient for human health and its global consumer demand is increasing. In order to promote the market supply and volumetric productivity of β-carotene using engineered Yarrowia. lipolytica strain. The fed-batch fermentation and DO-stat fed-batch fermentation were carried out. 73.5g/L biomass and 0.3g/L β-carotene were obtained in fed-batch. 94g/L biomass and 0.4g/L β-carotene were harvest during DO-stat fed-batch fermentation. The results of intracellular energy showed that more ATP and NADP+/NADPH were produced in DO-stat fed-batch fermentation as compared to that under fed-batch fermentation. Moreover, three kinetic models showed that the higher μm, Yx/s, and Yp/s values were achieved for DO-stat fed-batch fermentation. In this study, the DO-stat strategy was first reported for the β-carotene produce and the obtained results showed that it is a promising strategy in the industrialized β-carotene production.