Abstract: ENN41 is a freshwater strain isolated from Bo'Ao coast of Hainan Province. Microscopic observation showed that ENN41 has the unique morphological character of genus Botryococcus. The 18S rRNA gene of ENN41 was cloned for further species confirmation at molecular level. The homologous analysis showed that the 18S rDNA gene of ENN41 was highly homologous to the species of Botryococcus braunii in GenBank. ENN41 achieved a biomass productivity of 0.483 g/(Ld) and crude hydrocarbon content of 56.6% for 16 days culture in column reactor. The main fatty acids were oleic acid (C18:1), octadecatetraenoic acid (C18:4) and palmitic aid (C16:0), accounting for 72.6% of the total fatty acid. Large amount of lipid stained with Nile red in the extracellular matrix. ENN41 reached a high productivity after 16 days culture in plate bioreactor0.234 g/(Ld) with 20% crude hydrocarbon. All the results showed that ENN41 is a natural Botryococcus braunii strain with high productivity of biomass and lipids and high potential for industrial application.