作　　者： ; ; ; ;

机构地区： 天津科技大学生物工程学院

出　　处： 《粮油食品科技》 2012年第3期46-49,共4页

摘　　要： 实验以刺糖多孢菌CB11(Saccharopolyspora spinosa CB11)与红霉素高产菌株红色糖多孢菌E2(Saccharopolyspora erythraea E2)为亲本进行种间原生质体融合,通过对两亲本原生质体的不同灭活条件的考察,确定了S.spinosa CB11的热灭活条件为60℃水浴45 min,S.erythraea E2的紫外灭活条件为20 W紫外灯30 cm处,照射1 min。实验还优化了S.spinosa CB11与S.erythraea E2原生质体进行种间的最佳融合条件:室温下采用50%PEG 1 000融合3～5 min。融合子经多杀菌素的摇瓶发酵发现,融合子正突变率达到57.1%,其中遗传稳定性良好的高产融合子S.spinosa X9的多杀菌素产量可达290μg/mL,比亲本S.spinosa CB11(产量为153μg/mL)提高了89.5%。所有研究结果表明原生质体种间融合技术在刺糖多孢菌多杀菌素高产菌株选育中的应用是可行的、高效的。 The interspecies protoplast fusion between Saccharopolyspora spinosa CB11 and Saccharopolys- pora erythraea E2 were studied. The heatinactivated condition of S. spinosa CB11 and the UV inactivated condition of S. erythraea E2 were confirmed by test,which was for 45 min at 60℃ and 20 w UV lamp at a 30 cm vertical distance for 1 min. The optimal condition of protoplast fusion was that S. spinosa CBI I and S. erythraea E2 were co -incubated with 50% （w/w） PEG 1000 for 3 -5 rain at room temperature. The spinosad shake flask fermentation experiment showed that the positive mutation rate of fusants was 57.1% ,high spinosad - producing strain S. spinosa X9 was achieved, and the spinosad production was 290μg/mL, which was near to 89.5% higher than the initial strain S. spinosa CB11. The results showed that using protoplast fusion technique in the breeding of spinosad high - production strain of S. spinosa was feasible and efficient.

关 键 词： 原生质体融合 多杀菌素 刺糖多孢菌 红色糖多孢菌

领　　域： [轻工技术与工程] [轻工技术与工程]