Enzymes were an important role both in the food or beverage industry, but there is also an enzyme that acts as a breaker of the organism or cell wall of microorganisms. The enzyme known as biolytic enzyme. Extraction enzyme taken from the snail (Achatina fulica), especially in the abdomen, as in this section contains compounds glukorononidase β, and beta-glucanase and endo arylsulphatase (Ezeronye and Okerentugba, 2001). This enzyme was an important role before the fusion process takes place, namely for protoplast isolation. Furthermore, the enzyme used to break down the cell wall of the yeast Pichia manshurica. The yeasts are indigenus located around dahlia tubers and has the ability to produce inulinase (EC 18.104.22.168). Protoplast fusion is one way to improve the strain, so hopefully we will get a new strain and superior compared to its parent. The purpose of this research was using enzymes from snail to break the cell walls of yeast (protoplast isolation) and to obtain new fusan. Protoplast isolation performed enzymatically by using enzyme biolytic of snail. Fusion process was done by mixing the two parental pellets in solution osmotic stabilizer sorbitol solution of 1.0 mol / L containing 30% PEG 6000 (polyethylene glycol) and 10 mM CaCl2 for 20 minutes. The results showed that the concentration of biolytic 100% can be utilized as an agent biolytic enzyme and capable of producing protoplasts of 8.1 x 109 and it has been found that 3 fusan namely were Fusan D F1, F4 and F7 D.