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The freshwater microalga, Hydrodictyon reticulum, that contained 47.5% reducing sugars including 35% glucose was used as substrate for the production of l-lactic acid (LA) by LA-producing bacteria. Lactobacillus paracasei LA104 was selected for fermentation in a 5-l fermentor since it was able to grow at pH 3, 60g LA/l, 200g glucose/l, 125g NaCl/l, and 45°C and produced over 97.3% optically pure l-lactic acid with glucose as a substrate. Simultaneous saccharification and cofermentation from H. reticulum to l-LA using LA104 was investigated in a jar fermentor. The yield reached 46g/100g H. reticulum dry material, with a final concentration of 37.11g/l and a productivity of 1.03g/l/h. This is the first report of the production of l-LA from a microalga, and H. reticulum could be a potential feedstock for large-scale production of l-LA by LA104.