A new geometrial immobilization matrix with PVA fibers was introduced to enhance lactic acid production by immobilized Rhizopus oryzae. Compared to free cell fermentation, final lactic acid increased 25% (51.13g/L vs.40.9g/L) and fermentation time reduced 33% (48 h vs. 72 h) in shake-flasks. The immobilized-cell fermentation was evaluated for its long- term performance in both of shake-flasks and 1L bubble-column bioreactor operated in a repeated batch mode for 13 and nine cycles respectively. The overall yield in shake-flask was 65 %, corresponding to a volumetric productivity of 1.1 g/(L·h). Moreover, the highest lactic acid concentration in the bubble-column bioreactor was 55.68% with a yield of 69.6% from consumed glucose. This result suggested the promising use for the immobilization of Rhizopus oryzae in PVA fibers.