The effect of phosphatidylcholine on the molecular weight properties of hyaluronic acid (HA) was studied in batch culture of Streptococcus zooepidemicus by adding phosphatidylcholine at the early stage of exponential phase. With the addition of 80 mg/L of phosphatidylcholine, maximum HA yield (2.47 g/L) and weight-average molecular weight (902.60 KDa) were achieved, increased by 17.4% and 67.1%, respectively, as compared to the control. Metabolic flux analysis was employed to study the mechanism of phosphatidylcholine on the molecular weight of HA. The normalized flux distribution maps based on fermentation data at phosphatidylcholine addition indicated that phosphatidylcholine resulted in higher flux flowing to the HA pathway and lower flux flowing to the glycolysis and biomass synthesis pathway, coupling with higher level of UDPNAG generation and extra regeneration of ATP. The GC-MS analysis of fatty acids in the plasma membrane showed that the addition of phosphatidylcholine could promote the mobility and permeability of the cell membrane, making the HA chain pass through the membrane more easily, thus decreasing the energy consumption. All these results led to higher molecular weight of hyaluronic acid.