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

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。

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

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(UV-DES). 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.