Preparation of high yields of algal protoplasts using buccal juice of sea hare and commercial cellulase

Buccal juice of the sea hare Aplysia juliana was found to degrade algal pol ysaccharides. The optimal enzyme composition for protoplast preparation fro m Undaria pinnatifida was protein at 48 mu g/ml buccal juice from sea hare, 10 mg/ml cellulase Onozuka-RS, 0.4 M NaCl, 0.8 M sorbitol, 2 mg/ml dextran sulfate sodium salt, and 1 mu l/ml 2-mercaptoethanol in 10 mM MES buffer ( pH 6.0). Protoplasts of Eisenia bicyclis, Endarachne binghamiae (Phaeophyta ), and Ulva pertusa (Chlorophyta) could also be prepared in a similar manne r. Yields of these protoplasts were about 10(7) cells per gram of fresh wei ght alga.