Streptomycin resistance-aided genome shuffling to improve doramectin productivity of Streptomyces avermitilis NEAU1069
Streptomycin resistance-aided genome shuffling to improve doramectin productivity of Streptomyces avermitilis NEAU1069
复制标题
链霉素抗性辅助基因组改组提高阿维链霉菌 NEAU1069 的多拉菌素生产力
DOI：
10.1007/s10295-013-1280-8
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发表时间：
2013-05
影响因子：
3.4
通讯作者：
Xiang, Wensheng
中科院分区：
文献类型：
--
作者：
Zhang, Ji;Wang, Xiangjing;Diao, Jinna;He, Hairong;Zhang, Yuejing;Xiang, Wensheng
基因组改组是一种快速设计具有理想工业表型的微生物菌株的有效方法。为了快速提高阿维链霉菌NEAU1069的多拉菌素产量，本研究采用了链霉素抗性筛选与基因组改组相结合的方法。分别用N-甲基-N‘-硝基-N-亚硝基-N-亚硝基和紫外线(UV)处理出发菌株，筛选出5株高产多拉菌素的突变菌株作为GS-SR的起始菌株。最终获得了一株遗传稳定的菌株F4-137，摇瓶发酵产量为992±4.4 mg/L，分别是出发菌株UV-45和出发菌株NEAU1069的7.3倍和11.2倍。在50-L发酵罐中，F4-137菌株的多拉菌素产量达到930.3±3.8 mg/L，并对提高多拉菌素产量的相关因素进行了研究，结果表明，核糖体蛋白S12的突变和环己烷羧酸辅酶A产量的提高可能是改组菌株性能提高的原因之一。随机扩增多态性DNA分析表明，重组菌株之间存在遗传多样性，证实了基因组改组的发生。综上所述，我们的结果表明GS-SR是一种有效的提高链霉菌次生代谢产物产量的方法。
Genome shuffling is an efficient approach for the rapid engineering of microbial strains with desirable industrial phenotypes. In this study, a strategy of incorporating streptomycin resistance screening into genome shuffling (GS-SR) was applied for rapid improvement of doramectin production by Streptomyces avermitilis NEAU1069. The starting mutant population was generated through treatment of the spores with N-methyl-N’-nitro-N-nitrosoguanidine and ultraviolet (UV) irradiation, respectively, and five mutants with higher productivity of doramectin were selected as starting strains for GS-SR. Finally, a genetically stable strain F4-137 was obtained and characterized to be able to yield 992 ± 4.4 mg/l doramectin in a shake flask, which was 7.3-fold and 11.2-fold higher than that of the starting strain UV-45 and initial strain NEAU1069, respectively. The doramectin yield by F4-137 in a 50-l fermentor reached 930.3 ± 3.8 mg/l. Furthermore, the factors associated with the improved doramectin yield were investigated and the results suggested that mutations in ribosomal protein S12 and the enhanced production of cyclohexanecarboxylic coenzyme A may contribute to the improved performance of the shuffled strains. The random amplified polymorphic DNA analysis showed a genetic diversity among the shuffled strains, which confirmed the occurrence of genome shuffling. In conclusion, our results demonstrated that GS-SR is a powerful method for enhancing the production of secondary metabolites in Streptomyces.
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影响因子：
5
作者：
Ochi, Kozo;Hosaka, Takeshi
通讯作者：
Hosaka, Takeshi
影响因子：
2.6
作者：
Q. Jin;Zhi-hua Jin;Lijing Zhang;Shanjing Yao;Fuyong Li
通讯作者：
Q. Jin;Zhi-hua Jin;Lijing Zhang;Shanjing Yao;Fuyong Li
DOI：
--
发表时间：
2000
期刊：
--
影响因子：
--
作者：
T. Kieser
通讯作者：
T. Kieser
影响因子：
11.4
作者：
Wang, Pin-Mei;Zheng, Dao-Qiong;Wu, Xue-Chang
通讯作者：
Wu, Xue-Chang
影响因子：
4.4
作者：
Tanaka, Yukinori;Komatsu, Mamoru;Ochi, Kozo
通讯作者：
Ochi, Kozo