| Abstract |
This research was focused on investigation of the condition for protoplast formation and regeneration of protoplast fusion between Saccharomyces cerevisiae which has fermentation ability and Candida cariosilignicola which can grow at high temperature and utilize methanol. The results obtained were as follows; The highest production was collected in exponential growth phase. Ninety-nine% protoplast formation of C. cariosilignicola was obtained in glycin-NaOH buffer (pH10.0) containing Zymolyase 0.5mg/ml at 35℃ for 1hr incubation. The highest regeneration was produced when protoplast suspension containing 0.5% soft agar in buffered 50mM CaCl_2 was poured as a soft overlay onto 2% agar plates. Equal amount of protoplast suspension of two strains was mixed and centrifuged. The subsequent pellet was added to 2ml of 35% polyethylene glycol (MW 4,000) containing 50mM CaCl_2, and incubated at 30℃ for 10min. Then 0.1ml of the suspension of aggregated protoplast was immediately covered with minimal medium and incubated at 40℃ for 5-7 days. As results, SC_1, SC_2, and SC_3 fusants were obtained. The physiological characteristics of fusants produced by protoplast fusion were; SC_1, and SC_2 utilized maltose, galactose, methanol, potassium nitrate. SC_3 utilized all the above materials except galactose. |
