紫外-硫酸二乙酯复合诱变高通量筛选虫草素高产蛹虫草突变株

High-throughput screening of Cordyceps militaris for cordycepin production by UV-DES compound mutation

为了进一步提高虫草素的产量,本研究以蛹虫草(Cordyceps militaris)CICC 14014菌株为出发菌株,利用紫外-硫酸二乙酯(UV-DES)进行原生质体复合诱变,结合96孔板高通量筛选方法筛选虫草素高产且性状稳定的菌株。通过96孔板初筛获得54株虫草素产量大于6 g/L的突变株;再经三角瓶复筛,最终筛选出了一株虫草素高产突变菌株UD10-2,在液体表面培养条件下虫草素产量达到11.32 g/L,比发菌株CICC 14014(产量为5.43 g/L)提高了108.47%,经过20次传代后性状稳定。

In order to further improve the cordycepin yield,Cordyceps militaris（ C.militaris） CICC 14014 was considered as the starting strain for protoplast compound mutation.The protoplasts of the strain were induced by ultraviolet-diethyl sulfate（ UVDES）.Then,a high cordycepin yield and stable genetic strain was obtained by screening with 96-well microplates and triangle bottles.Through primary screening with 96-well microplates,54 mutant strains with a yield greater than 6 g/L cordycepin were obtained.A strain of cordycepin high yield mutant strain UD10-2 was screened out by secondary screenings with triangle bottles.The cordycepin production obtained by C.militaris UD10-2 mutant was 11.32 g/L in liquid surface culture,which was enhanced by 108.47% more than the original strain（ 5.43 g/L）.The results of subculture test showed that the mutant strain had stable hereditary characteristics after 20 generations.