Fed-batch cultures of Enterococcus faecalis RKY1 were performed to maximize the L(+)-Iactic acid concentration in the bioreactor. The highest lactic acid concentration was obtained at around 225 g/L by intermittent feeding the concentrated glucose media containing 500 g/L of glucose and 15 g/L (or 75 g/L) of yeast extract. However, in all fed-batch cultures, volumetric productivities of lactic acid gradually decreased due to the inhibitory effect of lactic acid produced during the fermentation. The highest value of lactic acid concentration obtained in this work corresponded to around 1.5-fold increase compared with conventional batch fermentation.