In this study, we focused on natural water-soluble antioxidants from fresh water microalgae, Pediastrum duplex and Dactylococcopsis fascicularis from Jeju Island, Korea. They were prepared by enzymatic digestion using five carbohydrases (Viscozyme, Celluclast, AMG, Termamyl and Ultraflo) and five proteases (Protamex, Kojizyme, Neutrase, Flavourzyme and Alcalase), and the potential antioxidant activity of each was assessed. All enzymatic digests from P. duplex showed significant DPPH scavenging effects. Termamyl (60.6%) digest from P. duplex possessed the highest effects on hydrogen peroxide scavenging. Celluclast (58.1%) and Kojizyme digests (56.9%) from D. fascicularis possessed higher effects on superoxide anion radical scavenging. All enzymatic digests exhibited significant effects on both NO· scavenging and metal chelating. Lipid peroxidation was significantly in inhibited Viscozyme, Termamyl and Kojizyme digests from P. duplex and Ultraflo, Protamex, Kojizyme and Alcalase digests from D. fascicularis. These data suggest that enzymatic digests of the fresh water microalgae, P. duplex and D. fascicularis might be valuable sources of antioxidant which can be applied in food and pharmaceutical industry.