酯化红曲霉原生质体制备及紫外线诱变育种

以红色红曲霉（Monascus ruber）为出发菌株,用混合酶制取原生质体并对原生质体形成条件进行探讨,并进行紫外诱变选育。采用50h菌龄的菌丝体,使用蜗牛酶（0.6%）＋溶菌酶（0.4%）＋纤维素酶（0.6%）的复合酶液,30℃酶解2h,在该条件下原生质体浓度可达7.8×106个/mL,再生率为7.3%。在最佳紫外照射时间80s下,突变株经筛选,得到一株酯化酶活显著提高、遗传性能稳定的诱变株HN215-6,其酶活达468.4mg/100mL,比出发菌株HN215提高了1.66倍。

Protoplast Preparation and Breeding of Higher Esterifying Enzyme Activity Mutants by Protoplast Mutagenesis of Ultraviolet Irradiation of Monascus Rubber

Using Monascus ruber as the original strain,Studied on the preparation and regeneration conditions of protoplasts by mutagenesis of ultraviolet irradiation.The results showed that treated 50h culture of Monascus ruber with 0.5% snail enzyme and 0.4% lysozyme and 0.6% Cellulose enzyme,the enzymolysis time was 2 hours,the enzymolysis temperature was 30℃,the protoplasts could reach 7.8×106 unit/mL,and the regeneration rate reached up to 7.3%.Under UV irradiation time in the best case for the 80s.After initially sieving and duplicate sieves,the mutagenesis strain which own increasing enzyme activity and transmissibility steady HN215-6 was obtained.Its esterifying enzyme content is 1.66 times higher than the original strain,Each 100 milliliter fermentation fluid esterifying enzyme activity achieves 468.4mg/100mL.