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《Liquor-Making Science & Technology》 2008-04
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Research on the Preparation and Regeneration of Protoplast from High-concentration Mash Resistant Saccharomyces cerevisia

YU Zhi-min, LUAN Jing, XU Peng, WANG Ji-hua and ZHAO Chang-xin (Liaoning Key Lab of Fermentation Technology, Dalian Polytechnic University, Dalian, Liaoning 116034, China)  
The preparation and regeneration conditions of protoplast from a high-concentration mash resistant S. cerevisia strain was studied. The results showed that the optimum time for protoplast preparation was the exponential growth of 7 h after the inoculum, and the optimum enzyme for cell wall hydrolysis was snailase. Through L16(5)4 orthogonal experiment, the optimum preparation conditions of S. cerevisia protoplast were identified as follows: 1.0 % of snailase, 0.7mol/L of KCl high osmotic buffer, 1.5 h hydrolysis, 0.1 % of β-mercaptoethanol protectant and hydrolysis temperature at 26 ℃. Under the above conditions, the formation rate and the regeneration rate of protoplast were 80.84 % and 36.03 % respectively. A further study showed that higher regeneration rate (39.78 %) of S. cerevisia protoplast could be obtained when the prepared protoplast was regenerated on the sandwich culture of 7 % sucrose media.
【Fund】: 辽宁省教育厅高校科研基金资助项目(NO2020701093)
【CateGory Index】: TS261.11
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