耐高糖酿酒酵母原生质体制备与再生过程研究  

Research on the Preparation and Regeneration of Protoplast from High-concentration Mash Resistant Saccharomyces cerevisia

作　　者：俞志敏[1] 栾静[1] 徐鹏[1] 王继花[1] 赵长新[1] 

机构地区：[1]大连工业大学辽宁省发酵工程重点实验室,辽宁大连116034

出　　处：《酿酒科技》2008年第4期45-48,共4页Liquor-Making Science & Technology

基　　金：辽宁省教育厅高校科研基金资助项目(NO2020701093)

摘　　要：对1株耐高糖酿酒酵母的原生质体制备和再生条件进行了研究。结果表明,发酵7h后为对数生长中期,适宜原生质体化;L1(65)确定制备原生质体的最佳条件为蜗牛酶浓度(1.0%)、KCl高渗缓冲液(0.7mol/L)、酶解时间(1.5h)、预处理剂(0.1%β-巯基乙醇)和酶解温度(26℃),在此条件下原生质体形成率和再生率分别为80.84%和36.03%;原生质体形成后在7%蔗糖高渗培养基上夹层培养再生率较高(39.78%)。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 ofS. 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 hy- drolysis temperature at 26 ℃. Under the above Conditions, the formation rate and the regeneration rate ofprotoplast 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.

关 键 词：微生物 耐高糖酿酒酵母 原生质体 制备 再生 

分 类 号：TS261.11[轻工技术与工程—发酵工程]