Abstract:86%。 By using single-factor test and orthogonal test, the optimized seed medium and feeding medium were obtained, with the aim to study the stability of the production of lactic acid by Rhizopus oryzae though semi-continuous fermentation. The optimization culture medium was as follows: glucose 120g/L,(NH4)2SO4 2g/L,NaH2PO4 0.16g/L,KH2PO4 0.14g/L,MgSO4?7H2O 0.2g/L,ZnSO4?7H2O 0.22g/L,CaCO3 60g/L; the feeding medium was as follows: glucose 100g/L,(NH4)2SO4 2g/L,KH2PO4 0.1g/L,ZnSO4?7H2O 0.33g/L,MgSO4?7H2O 0.15g/L,CaCO3 50g/L. In flask culture 30 repeated stable fermentation were carried out, and 20 batches were carried out in a 5 L stirred tank fermentor. The highest lactic acid yield from glucose was up to 86%.