非达霉素产生菌氮离子注入诱变及其发酵培养基优化

Breeding of fidaxomicin-producing strains by nitrogen ion implantation and optimization of the fermentation medium

目的通过氮离子注入对游动放线菌(Actinoplanes deccanensis)SIIA-A2098进行诱变、筛选高产突变株,并采用响应面法对突变株的发酵培养基进行优化,以提高非达霉素产量。方法采用多剂量氮离子注入对菌株SIIA-A2098进行诱变选育高产突变株。采用Plackett-Burman设计法筛选出影响突变株发酵的显著因素,然后采用CDD设计法分析优化显著影响因素参数。结果与结论氮离子注入得到非达霉素高产突变株SIIA M3510,产量为92.8μg/m L,是野生型菌株产量(11.5μg/m L)的8倍。Plackett-Burman设计法筛选出3个显著影响因素为葡萄糖、鱼粉蛋白胨和豆油,响应面法优化出它们的最佳数值分别为2.919%、0.644%和1.078%。在此优化条件下,摇瓶非达霉素发酵产量达到约244.75μg/m L,较优化前提高40.04%。

Objective High yield fidaxomicin mutants were screened from actinoplanes SIIA-A2098 based on N^＋ ion implantation technology. In order to greatly improve the fidaxomicin yield of the mutants, the consequent response surface methodology （RSM） was utilized to optimize the fermentation medium. Methods Multi dose nitrogen ion implantation were used to the strain SIIA-A2098 in order to breed high yield mutant. The Plackett- Burman design and response surface methodology were used to analyse and determine the important factors of the fermentation condition for producing fidaxomicin by the mutant. Results and conclusion Through the N^＋ ion implantation, we isolated a high-yield mutant SIIA-M3510, which yield productivity was eight-fold higher than the wild strain in the shake flask experiments. The glucose, fish peptone and bean oil were found to be the important factors and their optimized parameters is glucose 2.919%, fish pptone 0.644%, and bean oil 1.078%, respectively. The production of fidaxomicin could reach 244.75μg/mL by the optimal condition, which was increased by 40.04% compared with the original condition.