小诺霉素产生菌棘孢小单孢菌突变菌株原生质体形成、再生及融合的研究  

Studies on formation ,regeneration and fusion of protoplast in Micromonospora echinospora var.producer of micronomicin

作　　者：赵敏[1] 谢凤龙[1] 汪洁[1] 杨晓伟[1] 范瑾[1] 

机构地区：[1]江苏省微生物研究所

出　　处：《中国抗生素杂志》1995年第1期10-16,共7页Chinese Journal of Antibiotics

摘　　要：甘氨酸（Ｇｌｙ）的加入将会影响棘孢小单孢突变型菌株孢子发芽和菌丝生长，其影响程度与甘氨酸浓度及菌丝培养基成份有关，在Ⅱ号培养基中，０．３％Ｇｌｙ对菌丝形态的改变十分明显，而０．５％Ｇｌｙ能抑制孢子发芽。消色肽酶（ａｃｈｒｏｍｏｐｅｐｉｔｉｄａｓｅ）与溶菌酶（ｌｙｓｏｚｙｍｅ）联合作用比单一溶菌酶制备的原生质体形成率和再生率有明显提高。再生培养基以原分离培养基补充０．２ｍｏｌ／Ｌ蔗糖为宜，其再生频率可达到１５％。采用ＰＥＧ４０００，４２％（ｗ／ｖ）为助融剂，直接法检出融合重组体，其重组频率可达０．５％～１．０％，重组体中大部分苗株的次级代谢特性与亲株有较大差异，在再生菌落筛选中获得一些优良菌株，其小诺霉素产率较原亲株有显著提高。Sttidies on the formation of protoplast in M. echinospora var. shows that the presence of glycine(gly)could defer the germination of spora and grovvth of mycelia ofM.echinospora var.,the affection degree is dependent on the concentration of gly and the content of mycelium media,given medium Ⅱ 0.3 percent of gly could cause apparent changes in morphology of the mycelia,0.5 percent of gly almost compietly inhibit the germination of spora. Compared with lysozyme,the cofnbination of achromopeptidase andlysozyme could give mtich higher formation ratio of protoplast. The ratio of protoplastregeneration on separation media supplemented with 0.2mol/L sucrose could reach 15percent.The recombination ratio could reach 0.5～1.0 percent when 42%percent of PEG4000 (w /v)was used as promoter of the fusion and the recombinants were identified directly on dual drug plate,Many of the recombinants have different secondary metabolic characters with the mother strains. Some fine regenerators have much higher productivity of micronomicin than their mother strains.

关 键 词：小诺霉素 棘孢小单孢菌 原生质体 再生 融合 

分 类 号：Q939.5[生物学—微生物学] TQ465.2[化学工程—制药化工]