基于核糖体工程理论的常压室温等离子体诱变筛选多杀菌素高产菌  

Mutation Screening of Production Strains of Spinosad Based on Ribosome Engineering Theory through ARTP

作　　者：乔长晟[1,2] 赵男[1] 石漫漫 朱明[1] 李雪 

机构地区：[1]天津科技大学工业发酵微生物教育部重点实验室,天津300457 [2]天津北洋百川生物技术有限公司,天津300457

出　　处：《中国生物工程杂志》2014年第1期71-78,共8页China Biotechnology

摘　　要：利用核糖体工程理论,采用常压室温等离子体(ARTP)诱变对刺糖多孢菌(Saccharopolyspora spinosa)进行诱变筛选。以刺糖多孢菌QYLZ 88912菌株为出发菌株,对其进行ARTP诱变,选取三个不同致死率的照射时间,将孢子悬液混合,以增加抗性筛选几率。然后基于核糖体工程技术理论对混合孢子悬液进行初筛,最终筛选出了1株多杀菌素高产菌Sg200Rif110St40Er90-028。该菌株同时具有磺胺胍、利福平、链霉素、红霉素的多重抗性,摇瓶发酵试验表明,发酵7d后多杀菌素浓度可达到1 516.93 mg/L,较出发菌株QYLZ 88912的产量610.75 mg/L提高了148.37%,且遗传性稳定。以得到的高产菌Sg200Rif110St40Er90-028作为出发菌株,对发酵培养基进行响应面优化实验,优化后的培养基(g/L):葡萄糖46.97、麦芽糊精35、酵母粉40.36、水浸棉籽粉32.88、碳酸钙3,多杀菌素的产量为2 361.81 mg/L,比出发菌株的产量提高了286.71%。Based on the ribosome engineering theory, the atmospheric and room temperature plasma （ARTP） was used in screening of Saccharopolyspora spinosa. Taking the strain Saccharopolyspora spinosa QYLZ 88912 as the original strain, choose the spore suspension of three different irradiation time after ARTP mutation, then mix them to increase the resistance screening probability. A high-yield strain S. spinosa Sg200Rif110St40Er90-028 was obtained after screening to the mixed spore suspension. This strain had the resistance of sulfaguanidine, rifampin, streptomycin, erythromycin. Its production of spinosad reached 1516.93 mg/L under the level of shake flask fermentation through 7 days, which was 148.37% higher than that of the original strain S. spinosa QYLZ 88912. The subculture experiments indicated that the hereditary characteristic of high productivity of S. spinosa Sg200Rif110St40Er90-028 was relatively stable. The fermentation materials of spinosad producing strain were optimized by the response surface methodology based on the single factor experiment. Under the optimized medium, the spinosad was increased by 286.71% compared with the previous condition.

关 键 词：多杀菌素 核糖体工程 常压室温等离子体诱变 

分 类 号：Q52[生物学—生物化学]