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Abstract ▼ AI-Helper

In order to develope a protoplast fusion system for citric acid and SCP producing Candida lipolytica, the optimal conditions for the formation and regeneration of protoplast were examined and the protoplast fusion was performed. At the optimal conditions of growth phase and Zymolyase treatment, f...

In order to develope a protoplast fusion system for citric acid and SCP producing Candida lipolytica, the optimal conditions for the formation and regeneration of protoplast were examined and the protoplast fusion was performed. At the optimal conditions of growth phase and Zymolyase treatment, frequencies of protoplast formation were 98%. Approximately 20-30% of protoplasts were regenerated on the regeneration minimal medium containing 3% agar and 30mM $CaCl_2$ with the overlay of the same medium. The fusion frequencies, 4-5${\pm}$10$^{-4}$, were accomplished by the treatment of two nutritionally complementary auxotrophic protoplasts, L-14 ($lys^-$) and T-24 (X$30^-$), with 30% PEG 6000 containing 100mM $CaCl_2$ at $30^{\circ}C$ for 20 minutes.