筑波链霉菌诱变株的筛选与保藏方法研究

Breeding and Preservation Method of Mutant Strain of Streptomyces tsukubaensis

为了获得能稳定遗传的他克莫司生产菌种,供生产使用,本研究采用常压室温等离子体(ARTP)技术对他克莫司出发菌株TKM20-1进行诱变筛选,并对菌株保藏方法进行比较。结果表明:筛选获得1株可稳定遗传的突变高产菌株TKM21-15,相较于出发菌株,摇瓶效价提高了20%;对突变菌株进行4种保藏方式研究,确定该菌种可采用沙土管保藏法、冻干管保藏法、甘油管低温保藏法和甘油乳糖管保藏法来保藏,且沙土管保藏法和冻干管保藏法更适用于该菌种。

In order to obtain a stable genetic strain of tacrolimus for large-scale production,in this study,the atmospheric pressure room temperature plasma(ARTP)technology was used to mutagenize and select the tacrolimus starting strain TKM20-1,and strain preservation methods were compared.The results showed that a stable hereditary mutant high yielding strain TKM21-15 was obtained.Compared with the starting strain,the shake flask titer was obtained,increased by 20%.The mutant strain was studied in four preservation methods.It was determined that this strain can be preserved in sand soil tubes,freeze-dried tubes,glycerol cryogenic tubes and glycerol-lactose tubes.However,the sand tube preservation method and the freeze-drying tube preservation method are more suitable for this strain.