Potential feasibility of nutrients removal and biofuel production with microalgae was evaluated in batch culture. Distribution of microalgae in fresh water including reservoir and river was investigated to search for the species with high content of lipid that could converted into biofuel. Green algae, Chlorella and Scenedesmus sp., these are known as species containing high lipid content for biodiesel production, were observed in both summer and autumn season. However another highly lipid-containing species, botryococcus sp. was not observed in this study. In mixed culture of microalgae using synthesized wastewater medium, green algae were found to be dominant, comparing to other species of diatoms and blue-green algae. And microalgae were also capable of removing nitrogen and phosphorus in batch experiments. During the culture period of 14 days, removal efficiencies of nitrate and phosphorus were 30% and 82%, respectively. Furthermore, content of the intracellular lipid extracted from algae cell was as favorable as 12-30% in the mixed culture where Scenedesmus and Chlorella sp. were dominant. Therefore the mixed culture of microalgae could be applied to biofuel production and tertiary wastewater treatment, even though there are economic barriers to overcome.