质粒消除对冠菌素产生菌T1828生物学效应的影响

Effects of plasmid elimination on characteristics of coronatine-producing strain T1828

采用十二烷基硫酸钠(SDS)和提高生长温度结合法消除T1828菌株质粒后,筛选到无质粒突变株ZWL-15。以原始亲株T1828和无质粒突变株ZWL-15为出发菌株,考察了两者部分生物学特性。结果表明,菌株ZWL-15生长速率快于T1828,进入对数期和稳定期的时间分别提前1h和4 h。在菌株ZWL-15发酵过程中添加0.01%的SDS有利于冠菌素的合成,最适发酵温度比T1828升高5℃,达到35℃,发酵周期提前4 h,COR相对值是T1828的两倍左右。ZWL-15传代实验表明该突变株很稳定,且对氨苄青霉素敏感,氨苄青霉素基因初步定位于该菌属质粒上。

After eliminating plasmid of strain T1828 by combination sodium dodecyl sulfate（ SDS） with increased growth temperature, the plasmid-free mutant ZWL-15 was screened. The original parent strain T1828 and plasmid-free mutant ZWL-15 as parent strain, some biological characteristics of both strains were compared. The results showed that the growth rate of ZWL-15 was faster than T1828, and it was 1 h and 4 h earlier to get into the logarithmic phase and sta- tionary phase, respectively. Adding 0.01% SDS was useful for ZWL-15 to synthesizing coronatine in fermentation process When the optimum fermentation temperature 5℃ higher than that of the T1828, reached to 35℃, the fermentation cycle was 4 h earlier than that of the T1828, and the relative value of COR was about two times higher than that for T1828. Passage experiments of ZWL-15 indicated that the mutant ZWL-15 was very stable, and was sensitive to ampieillin. The ampieillin gene was targeted the plasmid in the genus.