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The biotechnological production of lactic acid could be carried out via anaerobic acidification process. In order to achieve an optimal production of lactic acid, the role of inoculum would be essential. The current study aimed to investigate as well as evaluate the effect of inoculum concentration on the anaerobic acidification of coconut water waste for the production of lactic acid. Results showed that the addition of 20% inoculums to the reactor fermenting coconut water waste was sufficient for the optimal production of lactic acid. In the batch process anaerobic acidification of coconut water waste inoculated with 20% inoculums of Lactobacillus acidophilus culture had the yield of lactic acid production, which was about 1.62 mmol lactic acid/mmol glucose while under the continuous operation the yield of lactic acid production obtained, was about 1.15 mmol lactic acid/mmol glucose. During the acidification process in both batch and continuous modes pH dropped significantly from 5.1 to 3.7.