不对称灭活原生质体融合技术选育川丁特罗高效转化菌株

Breeding of High Trantinterol Transforming Strains by Protoplast Fusion of Asymmetric Inactivation

分别以紫外线和加热灭活方法处理短刺小克银汉霉(Cunninghamellablakesleana)AS 3.970原生质体,并将2种方法灭活的原生质体在聚乙二醇(PEG)6000的作用下进行融合,从融合再生菌株中筛选川丁特罗高效转化菌株。结果显示,通过不对称灭活原生质体融合技术,获得1株川丁特罗高效转化菌株C. blakesleana AS 3.970 F-6,对川丁特罗的微生物转化率为(28.3±3.1)%,比原生质体融合前转化率[(8.2±2.6)%]提高了2.45倍。

The protoplasts of Cunninghamella blakesleana AS 3.970 were inactivated separately by UV irradiation and heat treatment,and then fused in polylethylene glycol(PEG)6000 for the selection of high trantinterol transforming strains.It was found that the UV death time on the protoplasts of C.blakesleana AS 3.970 was 9 min,and the heat treatment parameters were 55℃and 20 min.Eighteen fusion strains were obtained by protoplast fusion technology of asymmetric inactivation.A high trantinterol transforming strain F-6 was selected with the transformation rate of(28.3±3.1)%,which was 3.45 times higher than that of the original strain before protoplast fusion[(8.2±2.6)%].