1,3-Propanediol (1,3-PD) production is favored by high amount of glycerol which also contributes to the lethal 3-hydroxypropionaldehyde (3-HPA) accumulation. The influences of three culture conditions including initial glycerol concentration, stirring rate, and oxidoreduction potential (ORP) regulation on 3-HPA accumulation and 1,3-PD production were investigated. 3-HPA detoxification was available at initial glycerol concentration, aeration rate and stirring rate of up to 40gl −1 , 0.5vvm air flow and 250rpm, respectively. Alternatively, the stirring rate of 300rpm was sufficient to fulfill the fermentation with initial 50gglyceroll −1 . In addition, 3-HPA accumulation could be avoided at the initial 50gglyceroll −1 under the regulated reducing conditions (−200 and −400mV). However, ORP regulation was not convenient and feasible for the industrial production in that more carbon was distributed to weak organic acids. Consequently, two-stage fed-batch strategy was presented to achieve 3-HPA detoxification and maintain the expected high productivity of 1,3-PD, in which the batch culture stage was conducted at initial glycerol concentration and stirring rate of 40gl −1 and 250rpm, respectively, and the feeding culture stage was performed at stirring rate of 300rpm. 74.07g 1,3-PDl −1 was produced in 24h with the yield and productivity of 0.62molmol −1 and 3.08gl −1 h −1 .