阿卡波糖产生菌原生质体的制备与再生

目的研究阿卡波糖产生菌原生质体制备与再生的条件。方法通过溶菌酶破壁的方法制备原生质体,考察影响原生质体制备与再生的因素。结果和结论确认原生质体的制备条件为:一级培养采用SM2液体培养基,培养时间为33 h,转二级培养的转种体积分数为15%;二级培养采用R2YE培养基,培养时间为20 h,甘氨酸质量分数为0.7%;溶菌酶作用质量浓度为3 g.L-1,作用时间100 min,最大原生质体制备量达到8×1010个.L-1。考察了原生质体再生培养基的组成,在优化的再生培养基上,再生率达到9.2%。

Preparation and regeneration of protoplasts of Actioplanes sp., the producer of acarbose

Objective To investigate the conditions for the preparation and regeneration of protoplasts of Actioplanes sp.- the producer of acarbose. Methods protoplasts of Actioplanes sp. were prepared by bacteriolysis with lysozyme; the factors influencing the preparation and regeneration of protoplasts were investigated. Results and conclusions The conditions for the preparation of protoplasts were confirmed as following: SM2 medium was suitable for the first phase growth of Actioplanes sp; growing time was 33h; the inoculation volume of seed culture to second phase culture was 15 %（V/V）. Growing for 20h in R2YE medium was suitable for the second phase growth;. Glycine concentration in the R2YE medium was 0.7 %; 3mg/mL lysozyme concentration and 100 min digestion were suitable for lysis of cell wall. In above conditions the maximum amount of protoplasts prepared was 8×1010 L-1. Meanwhile, the regeneration medium was optimized; the regeneration rate of protoplasts was 9.2 % on the optimal regeneration medium.