啤酒酵母原生质体制备与再生条件的筛选

摘要

The conditions of preparation and regeneration of Saccharomyces cerevisiae Lnzenga protoplasts by orthogonal design were studied. Thernresult showed that the optimal preparing condition of S. Cerevisiae Lnzenga protoplast were as follows: mycelium static cultivation time 12h, additionrnof osmotic pressure stabilizer 0.6mol/L KC1, after pretreatment with EDTA-Na2+, using 2% snailase, enzymatic hydrolysis for 90min at 32℃. Underrnthese conditions, the preparation rate of protoplast achieved over 99%. Protoplasts produced under the conditions of mycelium static cultivation timern14h, addition of osmotic pressure stabilizer 0.6mol/L sucrose, no pretreatment, 1.5% snailase digesting for 120min at 30℃, the regeneration rate wasrn65%.%试验正交设计方法,对啤酒酵母原生质体制备与再生进行了研究.试验表明:以静置培养12h的细胞,0.6mol/L KCl作为渗透压稳定剂,经EDTA-Na2+预处理,在2％蜗牛酶作用下,32℃酶解90min为条件获得最佳制备率,制备率为99％以上；以静置培养14h的细胞,0.6mol/L蔗糖作为渗透压稳定剂,不经过预处理,在1.5％蜗牛酶作用下,30℃酶解120min,涂布于山梨醇为渗稳剂的再生培养基中为条件获得最佳再生率,再生率为65％.