In order to carry out the process of the repeated batch L-lactic acid fermentation by self-immobilized Rhizopus oryzae, we investigated the effect of medium compositions on the morphology of Rhizopus oryzae. In submerged culture, fungi can be grown as broths of freely suspended mycelia and pellets or clumps, the pellets can be immobilized by themselves. The optimum medium composition for the first patch of fermentation was: 120 g/L glucose, 3 g/L NH4NO3, 0.14 g/L KH2PO4, 0.16 g/L NaH2PO4 (the concentration ratio of K+ and Na+ was 1:1). After 72 h fermentation, the physical form of Rhizopus oryzae was mostly uniform pellet with the diameter of 1.0–2.0 mm , the concentration of L-lactic acid was 100.8 g/L, and the conversion rate of glucose 84%. During 16 batches of repeated fermentation, the L-lactic acid level was above 60.0 g/L and the glucose conversion rate was 75%. The time of every batch was 24 h.