【作者】 代鹏；

【导师】 林开春；

【作者基本信息】 华中农业大学 ， 农药学， 2004， 硕士

【摘要】 本文通过对多杀菌素产生菌刺糖多孢菌（Saccharopolyspora Jpinosa）9512-3菌株进行复壮，获得了一株发酵效价较高且稳定的菌株s-23。以s-23为菌株出发菌株，探索了用亚硝酸-氯化锂、紫外线、氮离子注入、⁶⁰Co-γ射线、超声波和紫外线复合5种物理、化学诱变方式对出发菌株进行诱变育种。结果发现：5种诱变作用后筛选到的菌株均能不同程度提高多杀菌的产素能力。其中紫外线、氮离子注入、⁶⁰Co-γ射线三种方式的诱变效果较好，分离到的三株高产菌uv-108-41、D-1-10及C121发酵效价分别比出发菌株效价提高13.8％、12.7％、12.6％。经过单孢分离纯化后的菌株C121发酵效价是原始菌株9512-3效价的420％。传代试验表明uv-108-41、D-1-10、C121三个菌株的遗传稳定性都比较稳定。 以MV-108-41菌株为试验菌株进行了发酵培养基的研究，探索了不同碳源，不同氮源和无机盐等对spinosad生物合成的影响。经L18（3⁷）两轮正交设计及七因子十三水平的均匀设计两个系统的筛选试验，最终筛选到两个发酵培养基的最佳配方。同时，对菌株uv-108-41的装料量、消前及消后pH、转速、温度等也做了研究发现：500毫升三角瓶中装料40ml；消前pH为7.0，消后6.7；转速210rpm：温度30℃条件下发酵效价最高达到386ug／ml。更多还原

【Abstract】 A stable mutated strain was obtained from the spores of spinosyn producer Saccharopolyspora spinosa 9512-3 by purification. The spores of s-23 were mutated by five methods Nitrous acid-LiCl, UV, 60Co irradiation, N+ transfusion, Cultrsaonic-UV. The result showed that five mutation methods could enhance the fermentation ability of strains. The chemical titer of the three high-yielding were increased by 13.8%, 12.7%,12.6% respectively compared with their parent strains. Subculture test indicated that uV-108-41, D-1-10, C121 had relatively stable hereditary characteristics.On shaking flask level, the fermentation media was examined with uv-108-41. Based on primary media, the effect of carbon,nitron,inorgonal to fermentation were investigated. We also screened the optional level of every component of the fermentation media by orthogonal and equality experiments. Two optimum fermentation medium were found.Experiment determining the sample contents, pH before sterilizing and after sterilizing, agitation rate, and temperature of uv-108-41 fermentation showed that the optional conditions were the volume 50ml/500ml, pH7.0 before sterilized, agitation 210rpm/min, temperature 30. The chemical titer can reach 386ug/ml.更多还原