摘要
使用纤维素酶和蜗牛酶复合酶液进行处理,并用β-巯基乙醇预处理,采用甘露醇作为渗透压稳定剂,考察菌龄、酶质量浓度、酶解温度和酶解时间对解脂亚罗酵母原生质体形成和再生的影响。实验得出结论:对于原生质体形成率影响因素来说,理论的最佳形成条件为:菌龄36 h,酶质量浓度为2.5mg/m L,酶解温度为25℃,酶解时间为8 h;对于原生质体再生率影响因素来说,理论的最佳再生条件为:菌龄48 h,酶质量浓度为2 mg/mL,酶解温度为25℃,酶解时间为8 h。而综合形成率和再生率共同影响因素顺序为:菌龄>酶质量浓度>酶解温度>酶解时间,最佳制备条件为:菌龄36 h,酶质量浓度2mg/mL,酶解温度25℃,酶解时间8 h。
The thallus of Yarrowia lipolytica was treated with cellulase and snail enzyme complex enzyme solution.It was pretreated with beta-mercaptoethanol and mannitol was used as the osmotic stabilizer.The effects of fungus age,enzyme concentration,enzymolysis temperature and enzymolysis time on protoplast formation and regeneration of Yarrowia lipolytic were investigated.It is concluded that the optimum conditions for protoplast formation were as follows:The fungus age was 36 h,the enzyme concentration was 2.5 mg/mL,the enzymolysis temperature was 25℃,and the enzymolysis time was 8 h.For the factors affecting protoplast regeneration,the best regeneration conditions were:The fungus age was 48 h,the enzyme concentration was 2 mg/mL,the enzymolysis temperature was 25℃,and the enzymolysis time was 8 h.And the order of the common factors affecting the formation rate and regeneration rate was:fungus age>enzyme concentration>enzymolysis temperature>enzymolysis time.The optimum preparation conditions were:the age of bacteria was 36 h,the enzyme concentration was 2 mg/mL,the enzymolysis temperature was 25℃,and the enzymolysis time was 8 h.
作者
侯晓娇
徐坤
HOU Xiao-jiao;XU Kun(School of Chemical Engineering,Eastern Liaoning University,Dandong 118003,China;Aimei Hissen Vaccine(Dalian)Co.Ltd,Dalian 116100,China)
出处
《辽东学院学报（自然科学版）》
CAS
2019年第1期41-47,共7页
Journal of Eastern Liaoning University:Natural Science Edition
关键词
解脂亚罗酵母
原生质体制备
原生质体再生
Yarrowia lipolytica
protoplast preparation
protoplast regeneration
