新西兰青霉(Penicillium novae-zeelandiae)原生质体的制备与再生研究

利用酶对新西兰青霉菌的菌丝进行酶解获得原生质体,探究了影响原生质体制备和再生的因素,包括酶种类,酶浓度,菌丝培养时间,酶解时间和温度,pH,二硫苏糖醇(DTT)预处理等.结果显示最佳酶解条件为:菌丝体培养48h,用0.05mol·L~(-1) DTT预处理0.5h,以0.8mol·L~(-1)氯化钠作为稳定剂,31℃条件下经Lywallzyme(10mg·mL~(-1))酶解4h,原生质体数量达到4.75×107 g~(-1),再生率为18.0%.

Formation and Regeneration of Penicilliumnovae zeelandiae HSD07B Protoplasts and Influence Factors

Protoplasts of Penicillium novae-zeelandiae HSD07 Bhave been successfully obtained by using lywallzyme on the mycelium.In order to achieve a high transformation frequency by producing these protoplasts,factors that affecting the preparation and regeneration were investigated which included the enzyme type,the enzyme concentration,the incubation time,the enzymolysis time,the enzymolysis temperature,pH and the preprocessing.After research,the maximum yield of protoplasts could be produced when mycelia was cultured for 48 h,pretreated by dithiothreitol(DTT)for 0.5h,and then digested for about 4hat 31 ℃in a buffer containing 10mg·mL~(-1) Lywallzyme.The amount of protoplasts could reach 4.75×107g~(-1) and the final regeneration rate was 18.0%.