地中海拟无枝酸菌原生质体电融合及其在提高利福霉素SV发酵效价中的应用  

Protoplasts electrofusion of Amycolatopsis mediterranei and its applications in improving the yield of rifamycin SV

作　　者：郑璞[1] 王蕾[1] 史朝辉[1] 

机构地区：[1]江苏省微生物研究所,无锡214063

出　　处：《中国抗生素杂志》2002年第5期267-269,279,共4页Chinese Journal of Antibiotics

基　　金：江苏省青年科技基金项目 (BQ980 4 6)

摘　　要：对产利福霉素SV的地中海拟无枝酸菌 (Amycolatopsismediterranei)原生质体进行热灭活和紫外灭活标记 ,其热灭活条件为 5 2℃ ,1.5h ,紫外灭活条件为紫外线照射 ( 15W ,2 5cm ,2 70nm) 6 0min。将双灭活标记的原生质体用CRY 3型细胞融合仪进行电融合 ,得出 :成串电流频率 1.5MHz ,电压 5 8V ,成串时间 2 0s ,融合脉冲幅宽 80 μS ,融合电压 5 0 0V ,融合槽间距 0 .5mm时 ,融合率最高为 9.3× 10 -4 。对筛选的融合子产量试验表明 ,5 0 %以上的融合子产量高于出发菌株 ,有明显的正变作用 ,将融合子在利福霉素代谢类似物平板上分离纯化 ,获得了化学效价提高 30 %以上的产量稳定菌株。The protoplasts of Amycolatopsis mediterranei were inactivated by heating 90min at 52℃ in water bath or ultraviolet irradiation 60min (15W, 25cm, 270nm). The inactivated protoplasts were electrofused in a CRY-3 Cell Fusion Apparatus . The optimal electronic parameters for orientating were as follows: 1.5 MHz and 58V ,action time 20s . After orientation, electronic pulses (500V, 80μs) were applied, the fusion rate was 9.3×10 -4 . The shake flask fermentation of the fusants showed that more than 50% fusants appeared higher yield than the control. After isolation with plate of PABA or ANA, two stable strains were obtained with yields of 30% higher than the control.

关 键 词：热灭活 紫外灭活 原生质体电融合 利福霉素SV 地中海拟无枝酸菌 

分 类 号：Q933[生物学—微生物学]