The use of different concentrations of sugar from sisal boles juice for production of Lactic acid (LA) using Lactic Acid Bacteria (LAB) has been studied with the intention of analysing the effect of initial sugar concentration, process temperature and initial medium pH on the produced lactic acid concentration, yield and productivity. All the linear variables (initial sugar conc., pH and temperature), two way and three way interactions were statistically significant for LA yield at p-values of less than 0.05 with correlation coefficient of 0.997. There was no significant effect on LA productivity of three way interactions (Temp.*pH*Initial Conc.). Maximum condition for production of LA occurred at a temperature of 37°C, initial pH of 6 and initial sugar concentration of 120 g/L which corresponded with the highest LA concentration of more than 24 g/Land a yield of 93%. This study shows that sisal boles juice has potential to produce LA.